01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-....

18
! !"#"$"%& "( !)* +,-./ 01!*-2,345 ,3 !)* !"#$%!&’(#)*+ ’*+(,!&)(-. +,-.67/ !&#* ,+ 5*0-*!,"3 5&5!*28 "9:; 7<;=><? ->@>AA; 2BCC9>DEFG? 2BAH;>9>>F 7EIA9>JJ K G? ;FC #;D<BAB; 0L M;N@<OPQB8 7>=;<DN>FD EJ #9;FD ;FC 2BA<E@B;9 .BE9E:O? 4FBR><PBDO EJ 0;9BJE<FB;? .><Q>9>O? 0S T/UVW? 45S ;FC K 7>=;<DN>FD EJ 0H>NBPD<O? 4FBR><PBDO EJ 0;9BJE<FB;? .><Q>9>O? 0;9BJE<FB; T/UVW? 45S ;FC #HOPBA;9 .BEPAB>FA>P 7BRBPBEF? $;X<>FA> .><Q>9>O 3;DBEF;9 $;@E<;DE<O? K 0OA9ED<EF -E;C? .><Q>9>O? 0;9BJE<FB; T/UVW? 45SL !"##$#% ’$’()* +$,-. ’/0/(/%1 233,)44 5/,,)40/#3)#5) ’/* 67’,$5$7 89 :7;<,14=$> ?)07,’;)#’ /@ 6(7#’ 7#3 A$5,/<$7( -$/(/%1> B#$C),4$’1 /@ 87($@/,#$7> -),=)()1> 82 D.EFG> BH2> I)(9 JKGLM.NLDFGN> OL;7$(* P7;<,14=Q#7’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‘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’$C) <75’),$7 "4) 40)5$7($P)3 4)5,)’/,1 ;)5S7#$4;4 ’/ ’,7#40/,’ ;75,/;/()5"()4 75,/44 ’S)$, )#C)(/0)4 ’/ ’S) )T’,75)(("(7, 4075) 7#3U/, ’/ ,)5$0$)#’ 5)((49 IS) )C/("’$/#7,$(1 ,)(7’)3 ’10) V+ 4)5,)’$/# 414’);4 WI.HH4X F ’,7#4@), ?Y2> 0,/’)$#4> /, </’S $#’/ /’S), <75’),$7 3",$#% 5/#Z"%7’$/#> ;73) )40)5$7((1 C$4$<() <1 ’S) 0,/07%7’$/# /@ 7#’$<$/’$5 ,)4$4’7#5)> /, ’/ )"=7,1/’$5 ,)5$0$)#’4 3",$#% <75’),$7( 07’S/%)#)4$49 I.HH4 7,) ,)["$,)3 @/, 07’S/%)#)4$4 <1 <75’),$7 ’S7’ 57"4) ;7#1 S";7# 3$4)74)4> 4"5S 74 \S//0$#% 5/"%S> <,"5)((/4$4> %74’,$5 "(5),4> 7#3 ])%$/##7$,)^4 3$4)74) W,)C$)\)3 $# WKX 7#3 WFXX9 R)#/;$5 4)[")#5) ;$#$#% ,)C)7(4 ;7#1 ;/,) I.HH4 /@ "#=#/\# @"#5’$/#9 !"#$%&’()#*+, (+,).&’*)/0> 7 4/$(L</,#) 0(7#’ 07’S/%)#> )#5/3)4 /#) /@ ’S) <)4’L "#3),4’//3 7#3 ;/4’ @()T$<() I.HH4 W,)C$)\)3 $# WNLJXX9 V# #7’",)> !"#$%&’()#*+, "4)4 ’S) 1*# L )#5/3)3 I.HH ’/ %)#)’$57((1 ;/3$@1 7 (7,%) #";<), /@ 3$5/’1()3/#/"4 0(7#’4 <1 ’,7#4@),,$#% 7 ?Y2 0/,’$/#> ’),;)3 ’S) IL4’,7#3> @,/; $’4)(@ ’/ 0(7#’ 5)((49 IS) 5)(("(7, 5S7#%)4 4)’ $# ;/’$/# <1 ’S) $#’)%,7’$/# /@ ’S) IL4’,7#3 %$C) ,$4) ’/ 5,/\# %7(( 3$4)74)9 V# ’S) (7</,7’/,1> ’S) 1*# L)#5/3)3 I.HH ’,7#4@),4 ?Y2 7#3 0,/’)$# 4"<4’,7’)4 ’/ <75’),$7> 0(7#’4> @"#%$> 7#3 ;7;;7($7# 5)((49 !"#$%&’()#*+, $4 5/;;/#(1 "4)3 ’/ 5,)7’) ’,7#4%)#$5 0(7#’4 @/, ,)4)7,5S 7#3 5/;;),5$7( 7%,$5"(’",)9 IS) ’\)(C) 0,/’)$#4 /@ ’S) 1*# L)#5/3)3 I.HH W,)C$)\)3 $# WWNLJXX 57# <) 3$C$3)3 $#’/ 4)C),7( 4"<%,/"04* KX ’S) ’,7#4@), 0$("4 WIL0$("4X> 5/;0/4)3 /@ +$,-F> ’S) ;7Z/, 0$($# 4"<"#$’_ +$,-J> 7 ;$#/, 0$($# 4"<"#$’_ 7#3 +$,-E 7 ($0/0,/’)$# ’S7’ ;71 S)(0 7#5S/, ’S) 0$("4 ’/ ’S) http://www.jbc.org/cgi/doi/10.1074/jbc.M606403200 The latest version is at JBC Papers in Press. Published on October 12, 2006 as Manuscript M606403200 Copyright 2006 by The American Society for Biochemistry and Molecular Biology, Inc. by guest on January 26, 2020 http://www.jbc.org/ Downloaded from

Upload: others

Post on 05-Jan-2020

10 views

Category:

Documents


0 download

TRANSCRIPT

Page 1: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

!

!"#"$"%&'"('!)*'+,-./'01!*-2,345',3'!)*'!"#$%!&'(#)*+

'*+(,!&)(-.'+,-.67/'!&#*',+'5*0-*!,"3'5&5!*28

"9:;'7<;=><?'->@>AA;'2BCC9>DEFG?'2BAH;>9>>F'7EIA9>JJKG?';FC'#;D<BAB;'0L'M;N@<OPQB8

7>=;<DN>FD'EJ'#9;FD';FC'2BA<E@B;9'.BE9E:O?'4FBR><PBDO'EJ'0;9BJE<FB;?'.><Q>9>O?'0S'T/UVW?

45S';FC'K7>=;<DN>FD'EJ'0H>NBPD<O?'4FBR><PBDO'EJ'0;9BJE<FB;?'.><Q>9>O?'0;9BJE<FB;'T/UVW?

45S';FC'#HOPBA;9'.BEPAB>FA>P'7BRBPBEF?'$;X<>FA>'.><Q>9>O'3;DBEF;9'$;@E<;DE<O?'K

0OA9ED<EF'-E;C?'.><Q>9>O?'0;9BJE<FB;'T/UVW?'45SL!"##$#%&'$'()*&+$,-.&'/0/(/%1

233,)44&5/,,)40/#3)#5)&'/*&67',$5$7&89&:7;<,14=$>&?)07,';)#'&/@&6(7#'&7#3&A$5,/<$7(&-$/(/%1>

B#$C),4$'1&/@&87($@/,#$7>&-),=)()1>&82&D.EFG>&BH2>&I)(9&JKGLM.NLDFGN>&OL;7$(*

P7;<,14=Q#7'",)9<),=)()19)3"

%<;N1F>:;DBR>' DO=>' ,+' P>A<>DBEF

POPD>NP' Y!/55PZ' D<;FPJ><'=<ED>BFP' ;FC'73S' DE

>IQ;<OEDBA' ;FC6E<' =<EQ;<OEDBA' <>AB=B>FDP

<>PI9DBF:' BF' =;DHE:>F>PBP' E<' AEF[I:;DBR>' 73S

D<;FPJ><L' +B<./?' EF>' EJ' DH>' NEPD' AEFP><R>C

=<ED>BFP' BF' DH>P>' POPD>NP?' H;P' @EDH' >F><:>DBA

;FC'PD<IADI<;9' <E9>P' BF' PI@PD<;D>' D<;FP9EA;DBEFL

\>'=<>RBEIP9O'=<>CBAD>C';'PD<IADI<;9'NEC>9'JE<

DH>'9;<:>'01D><NBF;9'CEN;BF'Y<>PBCI>P'/V]1U^TZ

EJ' +B<./' EJ' !/01234560789: 5896;3476<=L' )><>

X>' C>JBF>' ;' HENE9E:O1@;P>C' PD<IADI<;9' NEC>9

JE<'!/01234560789: +B<.KKL' .EDH' +B<./' ;FC

+B<.KK' NEC>9P' =<>CBAD' H>_;N><BA' E9B:EN><PL

&>;PD' DXE1HO@<BC' BFD><;ADBEFP' C>JBF>' =>=DBC>P

BF' DH>' 01D><NBFIP' EJ' +B<./' ;FC' +B<.KK' DH;D

BFD><;AD' XBDH' >;AH' EDH><L' !H>P>' BFD><;ADBEFP

X><>' N;==>C' EFDE' DH>' HENE9E:O' NEC>9P' DE

=<>CBAD' CB<>AD' BFD><;ADBEFP' @>DX>>F' DH>

H>_;N><BA'BFD><J;A>P'EJ'+B<./';FC'+B<.KK'PIAH

DH;D'DH>'+B<./'01D><NBFIP'PD;AQP';@ER>'+B<.KK

BF'DH>'=><B=9;PNL',F'PI==E<D?' J<;ADBEF;DBEF';FC

X>PD><F' @9EDDBF:' PHEX' DH;D' DH>' +B<./' 01

D><NBFIP' BP' 9EA;9B`>C' DE' DH>' N>N@<;F>' ;FC

=><B=9;PN?' <;DH><' DH;F' DH>' AODE=9;PN' EJ' A>99PL

SCCBDBEF;9' HB:H1<>PE9IDBEF' O>;PD' DXE1HO@<BC

<>PI9DP'C>NEFPD<;D>'BFD><;ADBEFP'@>DX>>F'DH>'01

D><NBFIP'EJ'+B<./';FC'DH>'=><B=9;PNBA'=E<DBEFP

EJ' +B<.K?' +B<.^?' ;FC' +B<.KWL' %>F>DBA' PDICB>P

<>R>;9'CENBF;FD'F>:;DBR>'BFD><;ADBEFP?';FC'DHIP

JIFADBEF?'EJ' DH>'+B<./'01D><NBFIP' 7<:>7>1L'!H>

;@ER>' C;D;' ;<>' BFD>:<;D>C' XBDH' DH>' >_BPDBF:

@ECO' EJ' 9BD><;DI<>' DE' =<E=EP>' ;' PD<IADI<;9?

=><B=9;PNBA' <E9>' JE<' DH>'01D><NBF;9'H;9J' EJ' DH>

!/01234560789:+B<./'=<ED>BFL

R,7;L#)%7'$C)& <75'),$7& "4)& 40)5$7($P)3

4)5,) '/ ,1 & ;)5S7# $ 4;4 & '/ & ' , 7#40/, '

;75,/;/()5"()4& 75,/44& 'S)$,& )#C)(/0)4& '/& 'S)

)T',75)(("(7,& 4075)& 7#3U/,& '/& ,)5$0$)#'& 5)((49& IS)

)C/("'$/#7,$(1& ,)(7')3& '10)& V+& 4)5,)'$/#& 414');4

WI.HH4XF& ',7#4@),& ?Y2>& 0,/')$#4>& /,& </'S& $#'/

/'S),&<75'),$7&3",$#%&5/#Z"%7'$/#>&;73)&)40)5$7((1

C$4$<()&<1&'S)&0,/07%7'$/#&/@&7#'$<$/'$5&,)4$4'7#5)>

/,& '/& )"=7,1/'$5& ,)5$0$)#'4& 3",$#%& <75'),$7(

07'S/%)#)4$49&I.HH4&7,)&,)["$,)3&@/,&07'S/%)#)4$4

<1&<75'),$7&'S7'&57"4)&;7#1&S";7#&3$4)74)4>&4"5S

74&\S//0$#%&5/"%S>&<,"5)((/4$4>&%74',$5&"(5),4>&7#3

])%$/##7$,)^4& 3$4)74)& W,)C$)\)3& $#& WKX& 7#3& WFXX9

R)#/;$5& 4)[")#5)& ;$#$#%& ,)C)7(4& ;7#1& ;/,)

I.HH4&/@&"#=#/\#&@"#5'$/#9

!"#$%&'()#*+,- (+,).&'*)/0>& 7& 4/$(L</,#)

0(7#'& 07'S/%)#>& )#5/3)4& /#)& /@& 'S)& <)4'L

"#3),4'//3&7#3&;/4'& @()T$<()&I.HH4& W,)C$)\)3& $#

WNLJXX9& V#& #7'",)>&!"#$%&'()#*+,& "4)4& 'S)& 1*#L

)#5/3)3& I.HH& '/& %)#)'$57((1& ;/3$@1& 7& (7,%)

#";<),&/@&3$5/'1()3/#/"4&0(7#'4&<1&',7#4@),,$#%&7

?Y2&0/,'$/#>& '),;)3& 'S)&IL4',7#3>& @,/;& $'4)(@& '/

0(7#'&5)((49&IS)&5)(("(7,&5S7#%)4&4)'& $#&;/'$/#&<1

'S)& $#')%,7'$/#&/@& 'S)&IL4',7#3&%$C)& ,$4)& '/& 5,/\#

%7((& 3$4)74)9& V#& 'S)& (7</,7'/,1>& 'S)& 1*#L)#5/3)3

I.HH& ',7#4@),4& ?Y2& 7#3& 0,/')$#& 4"<4',7')4& '/

<75'),$7>& 0(7#'4>& @"#%$>& 7#3& ;7;;7($7#& 5)((49

!"#$%&'()#*+, & $4& 5/;;/#(1& "4)3& '/& 5,)7')

',7#4%)#$5& 0(7#'4& @/,& ,)4)7,5S& 7#3& 5/;;),5$7(

7%,$5"('",)9

IS)& '\)(C)& 0,/')$#4& /@& 'S)& 1*#L)#5/3)3

I.HH& W,)C$)\)3& $#& WWNLJXX& 57#& <)& 3$C$3)3& $#'/

4)C),7(& 4"<%,/"04*& KX& 'S)& ',7#4@),& 0$("4& WIL0$("4X>

5/;0/4)3& /@& +$,-F>& 'S)& ;7Z/,& 0$($#& 4"<"#$'_

+$,-J>& 7& ;$#/,& 0$($#& 4"<"#$'_& 7#3& +$,-E& 7

($0/0,/')$#& 'S7'&;71& S)(0& 7#5S/,& 'S)& 0$("4& '/& 'S)

http://www.jbc.org/cgi/doi/10.1074/jbc.M606403200The latest version is at JBC Papers in Press. Published on October 12, 2006 as Manuscript M606403200

Copyright 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 2: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

!

!"#$%& '$'(%)*$+& ,-& #.$& #%)*/& $*0$1!2$& 2%!#$3*/

43%56& #.%!"7.&43%589+& :-& #.%$$& 3**$%;'$'(%)*$

)//!<3)#$=& 2%!#$3*/& >3#.& *"<1$!#3=$;(3*=3*7

=!')3*/?&43%@A?&43%5A?&)*=&43%588?&>.3<.&13B$1C

$*$%73D$& #%)*/2!%#$%& )//$'(1C& !%& /"(/#%)#$

#%)*/1!<)#3!*+& )*=& A-& 2%!#$3*/& >3#.& )==3#3!*)1

/2$<3E3<&!%&"*B*!>*&E"*<#3!*/&/"<.&)/F&)-&43%58?&)

#>!;=!')3*&2%!#$3*&>3#.& #>!&2%!2!/$=& E"*<#3!*/F

2$2#3=!71C<)*&1C/3/&#!&)//$'(1$&#.$&#%)*/&$*0$1!2$

<!%$& /#%"<#"%$?& )*=& 21)*#& 3*#$%)<#3!*& 03)& #.$

/$<%$#$=& 43%58& E%)7'$*#& 43%58G+& (-& 43%@A?

#.!"7.#&#!&<!"21$&#.$&@HI&/"(/#%)#$&#!&3#/&<!7*)#$

JAKK& #%)*/2!%#$%+& )*=& <-& 43%5:?& )& *$<$//)%C

2%!#$3*&!E&"*B*!>*&E"*<#3!*L

43%5A?& #.$& 1)%7$/#& JAKK& 2%!#$3*?& .)/& )#

1$)/#& #.%$$& 2%$=3<#$=& =!')3*/L& I*& H;#$%'3*)1

=!')3*& M%$/3="$/& 8N,;:NO-& ($1!*7/& #!& #.$

P)7QRJ%(QR43%5&STIU&E)'31C& M6-& )*=& !0$%1)2/

> 3 # . & ) & 2 " # ) # 3 0 $ & 4 3 % 5 A ;4 3 % 5 A

=3'$%3D)#3!*V'"1#3'$%3D)#3!*& =!')3*& M%$/3="$/& 8;

:8,-& MN-L& J.$/$& =!')3*/& )*=& #.$& )=W)<$*#& %$73!*

M%$/3="$/& :NO;A,O-& .)0$& *!& /3'31)%3#C& #!& B*!>*

/#%"<#"%$/&MX-L& Y*&<!*#%)/#?& #.$&P;#$%'3*)1&=!')3*?

>.3<.& <!*#)3*/& #.$& Z)1B$%& I& (![& M!%& S;1!!2-

M%$/3="$/& A::;AA9-& )*=& Z)1B$%& 5& (![& M%$/3="$/

68\;6:O-?&3/&2%$=3<#$=&#!&E!%'&)&.$[)'$%3<&J%>5;

13B$& /#%"<#"%$& MX-L& Y*=$$=?& (3!<.$'3<)1

E%)<#3!*)#3!*& %$0$)1/& #.)#&43%5A& E!%'/& !137!'$%/

!E&.$[)'$%3<&/3D$&M\-L&5!#.&#.$&43%588&)*=&43%@A

.!'!1!7/&)1/!&E!%'&.$[)'$%3<&!137!'$%/&M89?88-?

)&<!''!*&E$)#"%$&!E&IJS)/$/&M8,-&L

I11& #.%$$& IJS)/$/& M43%5A?& 43%588?

43%@A-& 2.C/3<)11C& 3*#$%)<#& )*=& )%$& 2%!2!/$=& #!

'$=3)#$& 3*3#3)1& J;/#%)*=& /"(/#%)#$& #%)*/2!%#& M8:-L

Z.31$&43%@A&)*=&43%588&.)0$&($$*&21)<$=&)#& #.$

$*#%)*<$& !E& #.$& JAKK& M8A-?& #.$& %$1)#30$& 2!/3#3!*

)*=&!%3$*#)#3!*&!E&43%5A&>3#.&%$7)%=&#!& #.$/$&#>!

.$[)'$%/?&)%$&*!#&B*!>*L&Z$&2%$03!"/1C&2%!2!/$=

#.)#&43%5A&>)/&1!<)#$=&3*&#.$&<C#!21)/'?&)<#3*7&3*

<!*<$%#& >3#.& 43%@A& MX-L& J.3/& )//37*'$*#& >)/

()/$=& /!1$1C& !*& #.$& /#%3B3*7& /#%"<#"%)1& /3'31)%3#C

($#>$$*& #.$& P;#$%'3*"/& !E& 43%5A& )*=& #.$

<C#!21)/'3<& =!')3*& !E&J%>5?& )&43%@A& .!'!1!7L

J.3/&.C2!#.$/3/&>)/&>3#.!"#&$[2$%3'$*#)1&/"22!%#L

]$%$?&>$& "/$& )& <!'(3*)#3!*& !E& C$)/#& #>!;.C(%3=?

(3!<.$'3<)1?& (3!3*E!%')#3<?& )*=& 7$*$#3<&'$#.!=/

#!& )==%$//& #.$& #!2!1!7C& !E& #.$& P;#$%'3*"/& !E

43%5AL

Z$& 2%$/$*#& #>!;.C(%3=& 3*#$%)<#3!*/

($#>$$*& #.$& P;#$%'3*"/& !E& 43%5A& )*=& #.$

2$%321)/'3<& =!')3*/& !E& /$0$%)1& 2%!#$3*/?& 43%58?

43%5X?& )*=& 43%589L& P$11& E%)<#3!*)#3!*& /#"=3$/

%$0$)1& #.)#& #.$& P;#$%'3*)1& .)1E& !E& 43%5A& 3/

1!<)13D$=& #!& #.$& 2$%321)/'?& )*=& 7$*$#3<& /#"=3$/

/"77$/#& 3#& 3/& 2.C/3<)11C& 3*0!10$=& 3*& #%)*/2!%#$%

E"*<#3!*& !"# $!$%L& Z$& E"%#.$%& !%3$*#& 43%5A& 3*

%$1)#3!*& #!&43%588L&J!&$*)(1$& #.$/$& 1)##$%& /#"=3$/?

>$&7$*$%)#$=&)&/#%"<#"%)1&'!=$1&!E&&'(%)*+,-(!./

43%588& ()/$=& !*& )& (3!3*E!%')#3</& <!'2)%3/!*

>3#.&#.$&/!10$=&/#%"<#"%$&!E&#.$&01#234%(!&.!'!1!7

]S9O,O& M89-?& )*=&2%$/$*#& )& %$E3*$=&'!=$1&!E&!"%

%$<$*#1C& 2%$=3<#$=& 43%5A& /#%"<#"%$& MX-L& 5C

<!'(3*3*7&!"%&E3*=3*7/&>3#.&$[3/#3*7&(3!<.$'3<)1

=)#)& M\?8:?8O-?& >$& /"77$/#& #.)#& #.$& P;#$%'3*)1

43%5A& .$[)'$%& %$/3=$/& 3*& #.$& 2$%321)/'& =3%$<#1C

)(!0$& )*& 3**$%& '$'(%)*$;$'($==$=& 43%588

.$[)'$%L

Q^SQ_YUQHJI`&S_aPQ@b_QK

54*6/!7# 8%"6,(.+,6# 9# c$)/#& #>!;.C(%3=& ()3#

E%)7'$*#/?&43%58& M))& X;,O,-?& 43%5A& M))& 8\X;A98

)*=&:,\;6O9-?&)*=&43%588&M))&8;896-?&>$%$&<1!*$=

3*#!& !"%& <$*#%!'$%$& ()/$=& C$)/#& #>!;.C(%3=

0$<#!%/?& 2P@L8& )*=& 2P@L,?& #.)#& 3*<%$)/$& #.$

/2$<3E3<3#C& !E& C$)/#& #>!;.C(%3=& 3*#$%)<#3!*/& M86-L

c$)/#& #>!;.C(%3=& 2%$C& E%)7'$*#/?& 43%58& M))& 68;

8A6-?&43%5A&M))&:,\;6O9?&A:,;O88?&A:,;O,,?&AA:;

O6A?&)*=&O,9;6O8-?&43%5X&M))&69;,:N-?&)*=&43%589

M))& 8OO;,A8-?&>$%$& %$#%3$0$=& E%!'& )&43%5& 13(%)%C

E"/$=& #!& #.$&d)1A& )<#30)#3!*&=!')3*& )<<!%=3*7& #!

#.$& '$#.!=/& =$/<%3($=& 3*& M8N-L& &'(%)*+,-(!./;

$[2%$//$=& %$73!*/& <!=3*7& E!%& 43%5A& M))& 8;NX\?

A:,;NX\?&)*=&AA:;NX\-&>$%$&<1!*$=&3*#!&2@Z9,\

>3#.& )*=& >3#.!"#& )*& H;#$%'3*)1& .$')71"#3*3*

M]I-&13*B$%L

2@Z9,\&>)/&<!*/#%"<#$=& #!&$*.)*<$&$!(;

%$7"1)#$=& 2%!#$3*& $[2%$//3!*& (C& E"/3*7& #.$& $!(:

2%!'!#$%& #!& )& <!*/$*/"/& %3(!/!'$;(3*=3*7& /3#$

M_5K-L&J.$&_5K&/$e"$*<$&)*=&#.$&;<%Y&)*=&84*Y

/3#$/& !E& 25S,8& M8X-&>$%$& %$21)<$=&>3#.& #.$&_5K

<!%%$/2!*=3*7& #!&&'(%)*+,-(!./# ,./-=*+!-"6&86K

%3(!/!')1& @HIL& Y*& #.$& 3*#%!="<$=& /$e"$*<$?

OfPJPdId>&>>&>>?JJdJP!"#IJPdIJ:f

?& #.$&_5K& 3/& 3*& 3#)13</& )*=& )&:620Y&/3#$&/2)**3*7

#.$& #%)*/1)#3!*& 3*3#3)#3!*& <!=!*& M(!1=-& 3/

"*=$%13*$=L&S%!#$3*&$[2%$//3!*&E%!'&#.3/&2%!'!#$%

3/&:;E!1=&.37.$%&%$1)#30$&#!&25S,8&)/&$/#3')#$=&(C

dTS& $[2%$//3!*& Mg"2)*?& Z)%=?& ])<B>!%#.?& )*=

g)'(%C/B3?&"*2"(13/.$=-L

@,(*!"6# *"7#8.4,.(-#8%"7!,!%"6&h&&'(%)*+,-(!./

,./-=*+!-"6& /#%)3*/& MPOX& M>31=;#C2$& *!2)13*$

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 3: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

!

!"#$%&'() $) *+,-.-/01) 234) 56#%7$"%76) 89:'() $&5

2;9<<=)8$)!"#$%)56>6"%?&')8:'')@6#6)A#?@&)?&)B;

89C) "#DE"?&6() <F3C) D6$!") 6G"#$H"() 9C) 0$2>') ?#

IJ;)8<F9C)D6$!")6G"#$H"()<F3C)K66L)6G"#$H"()<F3C

E6E"?&6() <F3C) !MH#?!6() NOP) PAQR=') @%"S) "S6

$EE#?E#%$"6) $&"%K%?"%H!F) Q"#$%&!) @6#6) A#?@&) $"

N4T2) $&5) $") 9:T2) "?) %&H#6$!6) U%#) E#?"6%&

6GE#6!!%?&F)&'()*#"()"+, (-.") !"#$%&!) 8/V3! ) $&5

56#%7$"%76!')@6#6) A#?@&) $") WXT2) ?&)B;)@%"S) "S6

$EE#?E#%$"6) $&"%K%?"%H!) 8Q%AO$'F) /+(()+#-01(*'

(*#*!"'"+*) !"#$%&!) 8I/99Y) 89Y') $&5) 56#%7$"%76!'

@6#6)A#?@&)$")N4T2)?&)IJ,/)89C)D6$!") 6G"#$H"(

NC)E6E"?&6()NC)A>MH?!6')?#)<FYXC)D6$!")&%"#?A6&

K$!6) @%"S?M") $O%&?) $H%5!) 8/%LH?) Z<:9:-93-W') "?

@S%HS) @6#6) $5565) $O%&?) $H%5) O%G6!) >$H[%&A

>6MH%&6() "#DE"?ES$&() $&5\?#) M#$H%>) $!) $EE#?E#%$"6F

1&"%K%?"%H) H?&H6&"#$"%?&!) 8%&) !A\OB') L?#) 2 3

450*6+("*7') @6#6]) [$&$ODH%&) 8N3'() "6"#$HDH>%&6

89<'() #%L$OE%H%&) 8N<') !E6H"%&?ODH%&) 89<<'() $&5

!"#6E"?ODH%&) 8W<<'F) 2?&H6&"#$"%?&!) L?#) &3, (-."

@6#6])H$#K6&%H%>>%&)89<<'()!E6H"%&?ODH%&)8X3'()$&5

!"#6E"?ODH%&)8N<'F

8#-4*"7, *9:#*''"-7)^)2;#-<+(4*#"+)A#?@&)?&)B;

$A$#) _\-) N<<!P) $H6"?!D#%&A?&6) @6#6) S$#76!"65(

#6!M!E6&565) %&) 56&$"M#%&A) !$OE>6) KMLL6#() $&5

K?%>65)L?#)3)O%&M"6!F),#?"6%&!)L#?O)9<4)H6>>!)@6#6

>?$565) %&"?) 6$HS) >$&6() #6!?>765) ?&)B$6OO>%) 8N<'

?#)"#%H%&6)8N9')E#?"6%&)A6>)!D!"6O!()$&5)"#$&!L6##65

"?) `OO?K%>?&-,)E?>D7%&D>%56&6)5%L>M?#%56)8,U/+'

<F=3!O) O6OK#$&6!) 8P%>>%E?#6) 2?#E?#$"%?&'F

Q"$&5$#5)E#?"?H?>!)L?#)a6!"6#&)K>?""%&A)@6#6)M!65

"?) 56"6H") U%#;=) 8!66) K6>?@) L?#) 56!H#%E"%?&) ?L

$&"%K?5%6!') ?#)V1-"$AA65)U%#;=) H?&!"#MH"!F)V1-

"$AA65) E#?"6%&!) @6#6) 56"6H"65) @%"S) O?&?H>?&$>

V1F99)82?7$&H6)b6!6$#HS)2?#E?#$"%?&')$&"%K?5DF

=#+(4"-7+4"-7)^)26>>!)@6#6) L#$H"%?&$"65)K$!65)?&

"S6)E#?H65M#6)?L)/6)P$$A5)$&5)BMA"6&K6#A)8NN'F

;#%6L>D() H6>>!)@6#6) S$#76!"65) $&5) #6!M!E6&565) %&

3<OP)E?"$!!%MO) ES?!ES$"6) KMLL6#() EV) 3F3() "S6&

E6>>6"65)KD)H6&"#%LMA$"%?&)$")X<<<)G);F)1&)$>%cM?"

?L) "?"$>) H6>>!) @$!) ?K"$%&65) $&5) L#?d6&) %&

56&$"M#%&A) !$OE>6) KMLL6#F) .S6) #6O$%&%&A) H6>>!

@6#6)#6!M!E6&565)%&)$&)?!O?"%H)!S?H[)!?>M"%?&)?L

3<OP) .#%!) EV) 4() N<C) !MH#?!6() NOP) J/.1(

4X!A\OB)ES6&D>O6"SD>!M>L?&D>) L>M?#%56) 8,PQ+'(

$&5) N<<!A\OB) >D!?dDO6) $&5) %&HMK$"65) W<

O%&M"6!) $") #??O) "6OE6#$"M#6F) .S6) O%G"M#6) @$!

H6&"#%LMA65) $") X<<<) G) ; ) $&5) "S6) !ME6#&$"$&"

8E6#%E>$!O%H)L#$H"%?&')@$!)H?&H6&"#$"65)KD)$H6"?&6

E#6H%E%"$"%?&F) .S6) E6>>6") @$!) #6!M!E6&565) %&

3<OP).#%!)EV)4)H?&"$%&%&A)4X!A\OB),PQ+) $&5

!?&%H$"65F) e&K#?[6&) H6>>!) @6#6) E6>>6"65) $&5

5%!H$#565F) .S6) O6OK#$&6) $&5) HD"?E>$!O%H

L#$H"%?&!)@6#6)!6E$#$"65)KD)"#6$"%&A)"S6)>D!$"6)@%"S

f2>) "?) $) L%&$>) H?&H6&"#$"%?&) ?L) <FNP) $&5

H6&"#%LMA$"%?&) $") NYN<<<) G) ;F) .S6) !ME6#&$"$&"

8HD"?E>$!O%H) L#$H"%?&') @$!) H?&H6&"#$"65) KD

$H6"?&6) E#6H%E%"$"%?&F) .S6) E6>>6") 8O6OK#$&6

L#$H"%?&') @$!) #6!M!E6&565) %&) $) KMLL6#) ?L) N<OP

.#%!) EV) XF=() 93<OP) f2>() 9<C) A>DH6#?>() $&5

<F<3OP) J/.1F) Q$OE>6!) @6#6) K?%>65) %&

56&$"M#%&A) KMLL6#) $&5) cM$&"%L%65) KD) KD) 7%!M$>

%&!E6H"%?&)?L)$)2??O$!!%6-!"$%&65)Q/Q-,1*J)A6>F

1EE#?G%O$"6>D) 6cM$>) $O?M&"!) ?L) E#?"6%&) @6#6

>?$565) %&) 6$HS) >$&6) ?L) $) 9<C)B$6OO>%) A6>) 8N<'(

#M&)$")N<<U)$&5)"#$&!L6##65)"?)$),U/+)O6OK#$&6F

;>?"!) @6#6) K>?H[65) %&) $) !?>M"%?&) ?L) 3C)O%>[) %&

.;Q.)$&5)"S6&)%&HMK$"65)@%"S)E#%O$#D)$&"%K?5%6!F

.S6) 0-"6#O%&$>) U%#;=) $&"%K?5D) %!) $) E?>DH>?&$>

#$KK%") $&"%K?5D) "?) "S6)0-"6#O%&$>)9<W)$O%&?)$H%5

#6!%5M6!) ?L) U%#;=) 8NW'F) .S6) 2-"6#O%&$>) U%#;=

$&"%K?5D) %!)$)E?>DH>?&$>) #$KK%")$&"%K?5D) "?) "S6)2-

"6#O%&$>) 93X) $O%&?) $H%5) #6!%5M6!) ?L) U%#;=) 8NW'

@S%HS)@$!)EM#%L%65)@%"S)$H6"?&6-E?@56#65)8N=')23

450*6+("*7') H6>>!) "S$")S$5)K66&)HM#65)?L) "S6%#).%

E>$!O%5F

>#+7'"*74, &9:#*''"-7, 2''+1) ^)? " ( - 4 " + 7 +

<*74)+0"+7+, E>$&"!) @6#6) A#66&S?M!6) A#?@&) L?#

L%76)"?)!676&)@66[!F)`&)$&D)?&6)6GE6#%O6&"()E>$&"!

L#?O) "S6) !$O6) H?S?#") @6#6) HS?!6&) @S6&) >6$76!

@6#6)$K?M")N)HO)%&)5%$O6"6#F)Q%&A>6)H?>?&%6!)?L)23

450*6+("*7')@6#6)A#?@&)L?#) "@?)5$D!)$")N4T2) %&

NOB) >%cM%5) B;) M&56#) !6>6H"%?&F) 9) OB) ?L) "S6

?76#&%AS")HM>"M#6!)@$!)E>$"65)?&"?)B;)E>$"6!)@%"S

$&"%K%?"%H!)$&5)A#?@&)L?#) "@?)5$D!)$")N4T2F)26>>!

@6#6) #6!M!E6&565) $&5) 5%>M"65) "?) $) L%&$>

H?&H6&"#$"%?&) ?L) R/Y<<) g) <FY) %&) $) !?>M"%?&

H?&"$%&%&A) 9<)OP)PA2>N() 9<)OP)PJQ) EV) 3F3(

$&5) N<<!P)$H6"?!D#%&A?&6F)26>>!)@6#6) %&HMK$"65

$")9:T2)L?#)"S#66)S?M#!)E#%?#)"?)%&L%>"#$"%?&F)Q%G)NY-

A$MA6) &665>6) S?>6!) @6#6) O$56) ?&) 6$HS, ?3

<*74)+0"+7+, >6$L) $&5) %&L%>"#$"65) @%"S) h=<<!B

!M!E6&!%?&)?L,23,450*6+("*7',E6#)>6$L)$!)56!H#%K65

89:'F) .S6) *+,-.-/01) E>$!O%5) @$!) $) A%L") L#?O

"S6) Q"$![$@%Hd) >$K() $&5) %!) $) 56#%7$"%76) ?L) EP/9

89:')O?5%L%65)"?)6GE#6!!)*+,)M&56#)"S6)H?&"#?>)?L

"S6)W3Q)E#?O?"6#F),>$&"!)@6#6)E>$H65)$")9:T2)L?#

9N)"?)N=)S?M#!)"S6&)"#$&!L6##65)"?)$)NNT2)%&HMK$"?#

@%"S)$)9Y)S?M#)>%AS")$&5)4)S?M#)5$#[)HDH>6F),>$&"!

@6#6)$!!$D65)L?#)*+,)6GE#6!!%?&)$")5%LL6#6&")"%O6!

5M#%&A)$)N=)"?)XN)S?M#)E6#%?5)E?!")%&L%>"#$"%?&F).6&

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 4: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

!

!"# $%# &'"()*+!&"',# -./.# .0+1&'.2# 3./# ("',!/)(!

3./# .03./&1.'!4# +'2#.03./&1.'!,#-./.# /.3.+!.2#5

!&1.,6

!"#$%&#%'() *+,) "-*./) '$%#/&&"+.#7#8*+'!# *.+9.,

-./.# 9&,)+*&:.2# "'# +# ;.&,,# <0&"3="!

.3&>*)"/.,(.'(.#1&(/",("3.#-&!=# +#?@8# >&*!./# ,.!

AB=/"1+# C.(='"*"DE# B"/3"/+!&"'4# ,.!# ')1F./

G$H$IJ6# K+*>L,.("'2# .03",)/.,# -./.# (+3!)/.2

!=/")D=# +# M"3./# N)+'!&0# (+1./+# FE# !=.# O8# P+F

3/"D/+1# AQ(+'+*E!&(,J6# C=.# ("*"/# !+F*.# +'2

F/&D=!'.,,# *.9.*,# -./.# +2R),!.2# &'# <2"F.

8="!",="3# &'# +# ("',&,!.'!# 1+''./# +(/",,# +**

.03",)/.,#+'2#.03./&1.'!,6

0%-%1%.()!%,/1"+.# 7#S.# ),.2# !=.#1"2.*# "># !=.

1"'"1./#">#!=.#BL!./1&'),#">#T&/UG#A%J#!"#(/.+!.

+'# &13/"9.2# 1"2.*# "># !=.# T&/UG# =.0+1./6# <,

3/.9&"),*E4# .+(=# ,)F)'&!# "># !=.# T&/UG# 1"'"1./

-+,# +*&D'.2# !"# !=.# ("//.,3"'2&'D# ,)F)'&!# "># !=.

,"*9.2# C/-U# =.0+1./# ,!/)(!)/.# A8/"!.&'# V+!+

U+'W# OV# $XYM# A$$JJ6#K"-.9./4# &',!.+2# "># ),&'D

!=.# 3/"D/+1#Z"2.**./# A[\J4# =./.#-.# ),.2# !=.# @&!

>)'(!&"'# "># V..3# T&.-# A[]J4# -=&(=# *.2# !"# +'

.13=+,&,# !"-+/2# /.,"*9.2# /.,&2).,# -=&*.# 2.L

.13=+,&:&'D#/.,&2).,#-&!=#'"#W'"-'#(""/2&'+!.,6

Z"2.*,# "># T&/UG# (+'# F.# +((.,,.2# +!

=!!3^__3=E*"D.'"1&(,6F./W.*.E6.2)_TOMUG6

C=.# T&/U$$# 3/"!.&'# ,.`).'(.# >/"1# 2 3

45-/6*#"/+&)-+,#+*&D'.2#-&!=# !=.#K8H\[\#3/"!.&'

,.`).'(.#>/"1#03)'(1%$")),&'D#!=.#+*&D'1.'!#D&9.'

&'#a."# /4) *16# [HHH# A$HJ6# C=.# Z"2.**./# 3/"D/+1

A[\J#-+,#),.2# !"#F)&*2#+#="1"*"DE#1"2.*#"># !-"

T&/U$$# ,)F)'&!,# F+,.2# "'# !=.# ,!/)(!)/.# "># !-"

,)F)'&!,#">#K8H\[\#>")'2# &'# !=.#+,E11.!/&(#)'&!

"># !=.#(/E,!+*# ,!/)(!)/.#">#K8H\[\#F")'2# !"#<V8

A8/"!.&'#V+!+#U+'W#b8VUc#OV#$?]de#(=+&',#<#+'2

U# A$HJJ6# C=.# =.0+1./# -+,# !=.'# +,,.1F*.2# FE

+33*E&'D# (/E,!+**"D/+3=&(# !=/..L>"*2# ,E11.!/E

"3./+!"/,#),&'D#!=.#3/"D/+1#d#A[IJ6

Z"*.()*+/# &1+D.,# -./.# 3/"2)(.2# ),&'D

!=.#fBQ@#B=&1./+#3+(W+D.#>/"1#!=.#M.,")/(.#>"/

U&"("13)!&'D4# T&,)+*&:+!&"'4# +'2# O'>"/1+!&(,# +!

!=.#f'&9./,&!E#">#B+*&>"/'&+4#Q+'#@/+'(&,("#A[%J6

MXQfPCQ

C=.# !"3"*"DE#"># !=.#BL!./1&'),#">#T&/UG

-+,# 2.>&'.2# ),&'D# ,.9./+*# .03./&1.'!+*

+33/"+(=.,6#S.#>&/,!#3/.,.'!# /.,)*!,#2.>&'&'D#,)FL

2"1+&',# "># 3/"!.&'L3/"!.&'# &'!./+(!&"',# F.!-..'

T&/UG# +'2# "!=./# ("13"'.'!,# "># !=.# CGQQ6# <,

2.,(/&F.2# F.*"-4# !=.# BL!./1&'),# "># T&/UG

&'!./+(!,# -&!=# 3./&3*+,1&(# 2"1+&',# "># ,.9./+*

("13"'.'!,#">#!=.#CGQQ6

78/) 9:4/$-"+*1) ,%-*"+) %6) ;"$<=) "+4/$*#4&) >"48

'/$"'1*&-"#)'$%4/"+&)?)C"#F.D&'#!"#+22/.,,#T&/UG

!"3"*"DE4#-.#2.>&'.2# &'!./+(!&"',#F.!-..'# !=.#BL

!./1&'),# "># T&/UG# A/.,&2).,# 5[YL]\HJ# +'2# "!=./

("13"'.'!,# "># !=.# CGQQ# ),&'D# !=.# E.+,!# !-"L

=EF/&2#,E,!.16#C=.#BL!./1&'+*#3"/!&"'#">#T&/UG#&,

"># 3+/!&()*+/# &'!./.,!# F.(+),.# &!# ("'!+&',# +# (*.+/

,!/)(!)/+*# 2"1+&'# A!=.# C/-UL*&W.# 2"1+&'J# -&!=

S+*W./# <# +'2# U# F"0.,# (=+/+(!./&,!&(# "># 1+'E

<C8+,.,6# @&D)/.# $# ,)11+/&:.,# T&/U$LT&/UG4

T&/U%LT&/UG4# T&/U$HLT&/UG4# +'2# T&/U$$LT&/UG

!-"L=EF/&2# &'!./+(!&"',6# S.# 3/.9&"),*E# /.3"/!.2

&'!./+(!&"',# F.!-..'# T&/UG# +'2# T&/U$4# T&/U%4

T&/U$H4#+'2#T&/U$$#A$IJe#="-.9./#A.0(.3!#>"/#!=.

T&/UGLT&/U%# &'!./+(!&"'J# !=.# 2+!+# 3/.,.'!.2# =./.

+/.# 2./&9.2# >/"1# '.-# !-"L=EF/&2# ,!)2&.,# !=+!

2.>&'.# ,1+**./# F+&!# +'2# 3/.E4# +'2# *.+2# !"# +# >&'./

1+3#">#!=.#/.D&"',#">#&'!./+(!&"'6

Q)33*.1.'!+/E# @&D)/.# $# 2.,(/&F.,# ")/

!-"L=EF/&2# +'+*E,.,# ),&'D# +# =&D=L/.,"*)!&"'# 3/.E

*&F/+/E# ("//.,3"'2&'D# !"# >),&"',# +>!./# .9./E

')(*."!&2.#">#!=.#@"$<#"3./"'#+'2#@"$A=6#C=.#=&D=

/.3/.,.'!+!&"'# "># !=.# *&F/+/E# 2.>&'.,# 3",&!&9.

&'!./+(!&"',#+D+&',!#+#=&D=*E#,&D'&>&(+'!#3"3)*+!&"'

"># '.D+!&9.# &'!./+(!&"',# +,# ("'!/"*,6# C=.# 2+!+# &'

@&D)/.# $<#2&,3*+E# /.D&"',# "># &'!./+(!&"'#F.!-..'

!-"# 3/"!.&',# +,# ,-+!=,# "># *+9.'2./# +'2# D/..'6

S=.'# !-"# &'2.3.'2.'!# &'!./+(!&"',# "9./*+34# !=.E

+/.# ,="-'#+,#2+/W./# ,-+!=,#"># ("*"/6#C=.,.# *+!!./

/.D&"',#">#"9./*+3#!=.'#2.>&'.#!=.#1&'&1+*#/.D&"',

">#&'!./+(!&"'#F.!-..'#!-"#3/"!.&',e#!=.,.#1&'&1+*

/.D&"',#+/.#,)11+/&:.2#&'#@&D)/.#$U6

T&/U$# =+,# F..'# 3/"3",.2# A[YJ# +'2# '"-

2.1"',!/+!.2# !"# =+9.# *E!&(# !/+',D*E(",E*+,.

+(!&9&!E#A5HJ4#!=+!#*&W.*E#>)'(!&"',#&'#!=.#3./&3*+,1

!"# (*.+9.# !=.# 3.3!&2"D*E(+'# *+E./# +**"-&'D

*"(+*&:.2#.,!+F*&,=1.'!#">#"!=./#3./&3*+,1&(#CGQQ

3/"!.&',6# O'# ,)33"/!4# $J# T&/U$# =+,# +# ,&D'+*

,.`).'(.#>"/# !/+',3"/!#+(/",,# !=.# &''./#1.1F/+'.

9&+#!=.#D.'./+*#,.(/.!"/E#3+!=-+E4#+'2#/.1"9+*#">

!=&,# ,.`).'(.# +F"*&,=.,# ("13*.1.'!+!&"'# "># +

@ " $ < B # 2.*.!&"'# 1)!+'!# A$%Je# [J# +*W+*&'.

3=",3=+!+,.#>),&"',#+*"'D#!=.#*.'D!=#">#!=.#T&/U$

("2&'D# ,.`).'(.# +/.# +(!&9.# A5$45[J4# +'2# 5J# !=.

T&/U$#BL!./1&'+*#3"/!&"'4#T&/U$g4# &,# .03"/!.2# !"

!=.# (.**# ,)/>+(.# +>!./# 3./&3*+,1&(# 3/"(.,,&'D

A$%455J6# 8/.9&"),# !-"L=EF/&2# 2+!+# ,="-#1)*!&3*.

T&/U$LT&/U%#+'2#T&/U$LT&/U$H#&'!./+(!&"',#A$IJ4

("',&,!.'!#-&!=# +#3./&3*+,1&(# *"(+*&:+!&"'# >"/# !=&,

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 5: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

!

!"#$%&"'( &)( '%&*+,-!.( /+%+( 0+( 1+2&-!*%3*+

%+4,'%&435( ,-*+%34*,&-!( #+*0++-(6,%7896,%7:( *;3*

!"''&%*('+%,'53!2,4(+<'&!"%+(&)(*;+(=9*+%2,-"!(&)

6,%7:(>?,$"%+(8@A(2,115+B.

C;+( ,-*+%34*,&-!( &)( *;+( =9*+%2,-"!( &)

6,%7:( 0,*;( 6,%7D( 3-1( 6,%78E( )"%*;+%( !*%&-$5F

!"$$+!*( *;3*( *;,!( %+$,&-( &)( *;+( 6,%7:( '%&*+,-

%+!,1+!( ,-( *;+( '+%,'53!2.( C;+( %+$,&-( &)( 6,%7D

,-*+%34*,&-( >%+!,1"+!(GE9HIJB( %+'%+!+-*!( *;+(+-*,%+

'+%,'53!2,4(1&23,-(&)(6,%7D(>KAI8AIHAI:9IGB.(L+

35!&( )&"-1( *;3*( 6,%7:( %+!,1"+!( 8KD9:E8( ,-*+%34*

0,*;( *;+(!32+('+%,'53!2,4(1&23,-(&)(6,%7D(>8JBM

*;"!A(*;+(2,-,235(6,%7D(,-*+%34*,&-(1&23,-(&44"%!

#+*0++-( 32,-&( 34,1!( IHK( 3-1( :E8( &)( 6,%7:

>?,$"%+( 8@A( *&'B.( C;&"$;( *;,!( %+$,&-( &)( 6,%7:

>%+!,1"+!( IHK9:E8B( ;3!( -&( N-&0-( !*%"4*"%+( &%

)"-4*,&-A( ,*!( ,-*+%34*,&-( 0,*;( 6,%7D( !"''&%*!( 3

'+%,'53!2,4( 5&435,O3*,&-( &)( *;,!( '&%*,&-( &)( *;+

6,%7:('%&*+,-.

P,N+0,!+A( 6,%7:( >%+!,1"+!( IHK9GQEB

,-*+%34*!( 0,*;( 6,%78E( >%+!,1"+!( 8QQ9H:8M( ?,$"%+

8@A(*&'BA(3(%+$,&-(*;3*(%+'%+!+-*!(3(53%$+('&%*,&-(&)

*;+(6,%78E('+%,'53!2,4(1&23,-( >%+!,1"+!(:J9IJJB

>KAI8AIJB.( C;"!A( #&*;( 6,%7:96,%7D( 3-1( 6,%7:9

6,%78E(,-*+%34*,&-!(!"''&%*( *;+(3!!,$-2+-*(&)( *;+

6,%7:(=9*+%2,-35(1&23,-(*&(*;+('+%,'53!2.

?"%*;+%( +R,1+-4+( )&%( 3( '+%,'53!2,4

5&43*,&-( &)( *;+( =9*+%2,-"!( &)( 6,%7:( ,!( ,*!

,-*+%34*,&-(0,*;(*;+(S9*+%2,-"!(&)(6,%788(>?,$"%+

8@A( #&**&2B.( C;+( &%,+-*3*,&-( &)( *;+( !&5R+1

!*%"4*"%+( &)( *;+( 6,%788( ;&2&5&$( /TEQHQ( )%&2

!"#$%&'(%)"*+ ,-#&*$( >8EB( '%+1,4*!( *;3*( 6,%788

%+!,1+!(,-(*;+(,--+%(2+2#%3-+(0,*;(,*!(S9*+%2,-"!

)34,-$(*;+('+%,'53!2(3-1(,*!(=9*+%2,-"!()34,-$(*;+

4F*&'53!2.( C;+( ./*&'(%)"*$01( 6,%78896,%7:

,-*+%34*,&-(,!(3-35FO+1(,-(1+*3,5(#+5&0.

!&1&#&/-+ 1&2"#+ &3+ 4$*566+ '(7"2+ &8+ !9+ ,-#&*$

!:;<=<(U(C&(*+!*(0;+*;+%(*0&9;F#%,1(13*3(43-(#+

"!+1( *&( 1+*+%2,-+( ;&0( )"5595+-$*;(C:VV( '%&*+,-!

,-*+%34*( ,-( *;+( 4+55A( 0+( 3-35FO+( *0&9;F#%,1

,-*+%34*,-$(1&23,-!( 3-1( !*%"4*"%35( 13*3( *&( '%+1,4*

*;+( *;%++(1,2+-!,&-35( %+53*,&-!;,'(#+*0++-(6,%7:

3-1( 6,%788.( V,-4+( *;+( !*%"4*"%+!( &)( 6,%7:( 3-1

6,%788( 3%+( "-N-&0-A( 0+( ),%!*( -++1+1( *&( #",51

;&2&5&$F(2&1+5!(&)(*;+!+('%&*+,-!.(@(!*%"4*"%+(&)

6,%788( 03!( '%+1,4*+1( "!,-$( #,&,-)&%23*,4!

3''%&34;+!( !,2,53%( *&( *;&!+( "*,5,O+1( *&( 4%+3*+( 3

;&2&5&$F(2&1+5(&)(*;+(6,%7:(=9*+%2,-"!(>DB.

W"%(6,%788(;&2&5&$F(2&1+5( ,!(#3!+1(&-

/TEQHQ()%&2(!9+,-#&*$>(0;,4;(!;3%+!(I8X(32,-&

34,1(,1+-*,*F(>3-1(QH(X(!,2,53%,*F(&R+%(IE8(35,$-+1

%+!,1"+!B( 0,*;( .9+ )01"3(%$"87( 6,%788.( C0&

!"#"-,*!(&)(/TEQHQ(0+%+()&"-1(,-(*;+(3!F22+*%,4

"-,*(&)(,*!(4%F!*35(!*%"4*"%+(>8EB(3-1()&%2(*;+(#3!,!

)&%( &"%( !*%"4*"%35( 4&2'3%,!&-(0,*;( 6,%788.( C0&

2&-&2+%!( &)( &"%( '%+1,4*+1( 6,%788( !*%"4*"%+

>?,$"%+( H@A( %,$;*B( 3%+( ;,$;5F( !,2,53%( *&( *0&

2&-&2+%!(&)(/TEQHQ(>?,$"%+(H@A(5+)*B.(=&2'3%+1

*&(&"%(6,%7:(2&1+5A(*;+(6,%788(2&1+5(03!(+3!,+%

*&(4&-!*%"4*(1"+( *&( *;+(;,$;+%(1+$%++(&)( !+Y"+-4+

,1+-*,*F( 3-1( !,2,53%,*F( #+*0++-( 6,%788( 3-1

/TEQHQ.(73!+1(&-(*;+(%+53*+1-+!!(,-(!+Y"+-4+(3-1

)"-4*,&-( 0,*;( /TEQHQA( 6,%788( ,!( '%+1,4*+1( *&

)&%2( 3( ;+<32+%( >?,$"%+( H7B.( Z)(6,%788( 31&'*!( 3

!,2,53%( 2+2#%3-+( &%,+-*3*,&-( 3!( /TEQHQ( >8EBA

*;+-(,*!(S9*+%2,-35(1&23,-(0&"51(#+(+2#+11+1(,-

*;+(,--+%(2+2#%3-+()34,-$(*;+('+%,'53!2(>TBA(0,*;

,*!( =9*+%2,-35( 1&23,-( +<'&!+1( *&( *;+( 4F*&'53!2

>=B(>?,$"%+(H=A(#&**&2(5+)*B.

Z-(311,*,&-( *&(&"%(-+0(6,%788(2&1+5A(0+

,2'%&R+1( &"%( ;+<32+%,4( 2&1+5( &)( *;+( 6,%7:( =9

*+%2,-"!( >%+!,1"+!(:HQ9JDKB( >DB.( C;,!( -+0(2&1+5

+2';3!,O+!( %+!&5R+1( %+!,1"+!( 5+31,-$( *&( 3( 2&%+

4&2'34*( 3-1( !F22+*%,435( *;%++91,2+-!,&-35

!*%"4*"%+(>?,$"%+(H=A("''+%(%,$;*(3-1(5+)*(,23$+!B.

?(%"7+ &3+ 4$*5@+ (82+ 4$*566+ A"B(1"*7+ $8)"*(%)(U

L+( 23''+1( 6,%7:96,%788( *0&9;F#%,1

,-*+%34*,&-!(&-*&( *;+( *;%++(1,2+-!,&-35(2&1+5!(&)

6,%7:( 3-1(6,%788( *&( 3!!+!!( 0;+*;+%( *0&9;F#%,1

,-*+%34*,&-!(,-45"1+(%+!,1"+!(*;3*(3%+(344+!!,#5+()&%

'%&*+,-9'%&*+,-(,-*+%34*,&-.(Z-(*;+(F+3!*(*0&9;F#%,1

3!!3FA( *;+( ),%!*( *;,%1( &)( 6,%788( >%+!,1"+!( 898EGB

%+*%,+R+1( 3( %+53*,R+5F( !2355( %+$,&-( &)( 6,%7:

>%+!,1"+!( ::I9QG:B( >?,$"%+( 8@A( #&**&2B.( C;+!+

1&23,-!( &)( ,-*+%34*,&-( #+*0++-( 6,%7:( >%+!,1"+!

::I9QG:B( 3-1( 6,%788( >%+!,1"+!( 898EGB( 0+%+

23''+1( ,-( *+35( 3-1( #5"+A( %+!'+4*,R+5FA( &-*&( *;+

%,##&-( >!,1+( R,+0B( 3-1( !'34+( ),55,-$( !*%"4*"%35

2&1+5!( &)( 6,%7:( 3-1( 6,%788( >?,$"%+( H=B.

V*%,N,-$5FA( *;+( %+$,&-!( &)( ,-*+%34*,&-( #+*0++-

6,%7:(>*&'A( ,-( *+35B(3-1(6,%788(>#&**&2A( ,-(#5"+B

23'( *&( 3( !,-$5+( )34+( &)( +34;( ;+<32+%.( [-&0,-$

*;+( &%,+-*3*,&-( &)(6,%788( ,-( *;+( ,--+%(2+2#%3-+

355&0!( "!( *&( &%,+-*( 6,%7:.( 7F( 3-35&$F( *&( *;+

*&'&5&$F(/TEQHQ( >8EB( *;+(S9*+%2,-"!( &)(6,%788

)34+!( *;+( '+%,'53!2( >*&03%1!( *;+( +<*+%,&%( &)( *;+

4+55B.( C;+%+)&%+A( *;+( =9*+%2,-"!( &)( 6,%7:( 2"!*

%+!,1+( ,-( *;+( '+%,'53!2( 0,*;( *;+( !"%)34+( *;3*

,-*+%34*!(0,*;(*;+(S9*+%2,-"!(&)(6,%788()34,-$(*;+

,-*+%,&%(&)( *;+( 4+55.(C;+!+( %+!"5*!(1+2&-!*%3*+( *;+

4&2'5+2+-*3%F("*,5,*F(&)(*0&9;F#%,1(3-1(!*%"4*"%35

13*3.

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 6: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

!

!"#$%& '()$*& *+,-"'./01& 02*(/)3*0,2$

.(*+((2&40/56&)21&7(/078)$903&!6::&3,97,2(2*$

;40/5<%&40/5=%&40/5<>?& 3,9.02(1&+0*"&9)7702@

,A& 40/56-40/5<<& 02*(/)3*0,2& 1,9)02$& ,2*,

$*/#3*#/)8& 9,1(8$& ,A& 40/56& )21& 40/5<<

3,2$0$*(2*8'& $#77,/*& )&7(/078)$903& 8,3)*0,2&,A& *"(

B-*(/902#$& ,A&40/56C&50,3"(903)8& $*#10($& .(8,+

3,2A0/9&*"0$&)$$0@29(2*C

!"#$%& '()*#+",-.& /#-0+*,)& "1& /23,4& ",& )5*

6*#"6.-1+&D&!,&)11/($$&10/(3*8'&+"(/(& 02& *"(&3(88

*"(& B-*(/902#$& ,A& 40/56& 7/,*(02& 0$& 8,3)*(1%& 3(88

A/)3*0,2)*0,2&)21&E($*(/2&.8,**02@&+(/(&7(/A,/9(1

,2&70#28-9)*#"3+& 3(88$& (F7/($$02@&.,*"& *"(&+081-

*'7(& 40/56& )21& )& "(9)@8#*0202& ;GH?-*)@@(1

40/56& B-*(/902#$& ;/($01#($& 6IJ-K=L?C& E(

9,20*,/(1& A/)3*0,2$& #$02@& )2*0.,10($& *,& *"/((

10AA(/(2*& 7(7*01($%& 40/56& M-*(/902)8& /($01#($& <-

<>I%&40/56&B-*(/902)8& /($01#($& NIJ-K=L%& )21& *"(

GH& (70*,7(& ;O0@#/(& I?C& !"(&M-*(/902)8& )2*0.,1'

1(*(3*(1&*"(&A#88-8(2@*"&40/56&7/,*(02&02&*,*)8&3(88$

)21&02&*"(&9(9./)2(&A/)3*0,2&;O0@#/(&IH?C&!"(&B-

*(/902)8&)2*0.,1'%&",+(P(/%&1(*(3*(1&*+,&10AA(/(2*

40/56&$7(30($&+",$(&$0Q($&3,//($7,21&*,& *"(&A#88-

8(2@*"&7/,*(02&)21&*"(&B-*(/902)8&A/)@9(2*&;O0@#/(

I5?C&!"(&A#88-8(2@*"&7/,*(02&)@)02&+)$&1(*(3*(1&02

*"(& *,*)8& 3(88$& )21& 02& *"(& 9(9./)2(& A/)3*0,2

;O0@#/(& I5%& #77(/& )//,+"()1$?C& !"(& B-*(/902)8

A/)@9(2*&+)$&1(*(3*(1&02&*"(&*,*)8&3(88$&)21&02&*"(

9(9./)2(& A/)3*0,2& )$& +(88%& .#*& #280R(& *"(& A#88-

8(2@*"&7/,*(02%& 0*&+)$& A,#21&)8$,& 02& *"(&7(/078)$9

A/)3*0,2& ;O0@#/(& I5%& 8,+(/& )//,+"()1$?C& !"(&GH

)2*0.,1'& 3,2A0/9(1& *"0$& 8)**(/& /($#8*& .'& )@)02

1(*(3*02@& *"(& B-*(/902)8&40/56& A/)@9(2*& 02& *,*)8

3(88$& )$& +(88& )$& 02& *"(& 7(/078)$9& )21& 9(9./)2(

A/)3*0,2$& ;O0@#/(& IB?C&E(& 2(P(/& ,.$(/P(1&40/56

;,/& 0*$& B-*(/902)8& A/)@9(2*?& 02& *"(& 3'*,78)$9

A/)3*0,2& #21(/& )2'& 3,210*0,2$C& S2& 3,2*/,8

(F7(/09(2*$%&40/5L%&)&3,/(&7(/078)$903&7/,*(02&,A

*"(&!6::%&+)$&A,#21&02&*,*)8&3(88$%&7(/078)$9%&)21

9(9./)2(& A/)3*0,2$& ;O0@#/(&IT?%&+"08(& *"(&!6::

$#.$*/)*(&7/,*(02&40/UJ&+)$&1(*(3*(1&02&*,*)8&3(88$%

3'*,78)$9%& )21& 9(9./)2(& A/)3*0,2$& ;1)*)& 2,*

$",+2?C&!)R(2&*,@(*"(/%&*"($(&/($#8*$&02103)*(&*")*

*"(& A#88-8(2@*"& 40/56& 7/,*(02& 0$& )23",/(1& 02& *"(

9(9./)2(%& 7,$$0.8'& .'& 0*$& M-*(/902)8& 1,9)02%

+"08(& *"(& B-*(/902)8& 1,9)02& 0$& 8,3)*(1& 02& *"(

7(/078)$9C

:#-0+*,)1& 2/& !"#$%& *;*#)& -& 42+",-,)& ,*0-)"<*

*//*9)& 2,&=(>?7& )#-,1/*#&D&H& A/)@9(2*& ,A&40/56

$*)/*02@&)*&/($01#(&6IJ&1(A02(1&*"(&$9)88($*&/(@0,2

,A&40/56& 02*(/)3*0,2&+0*"& 0*$(8A& ;1)*)& 2,*& $",+2?

;<K?&)21&+0*"&40/5<%&+"08(&)& A/)@9(2*&$*)/*02@&)*

/($01#(& 66I& +)$& *"(& $9)88($*& /(@0,2& 02*(/)3*02@

+0*"& 40/5<<& ;O0@#/(& <?C& :0@20A03)2*8'%& *"(

3)2,203)8& E)8R(/& H& .,F& $0*$& 7/(30$(8'& .(*+((2

)902,&)301$&6IJ&)21&66I%&$#@@($*02@&*")*&*"(/(&0$&)

10$*023*& 10AA(/(2*0)*0,2& 02& !6::& 3,97,2(2*

02*(/)3*0,2& .(*+((2& *"(& /(@0,2& B-*(/902)8& ,A& *"(

E)8R(/& H& .,F%& )21& ,2(& 0238#102@& *"(&E)8R(/& H

.,FC&E0*"&*"0$&02&9021%&+(&)$$)'(1&*"(&A#23*0,2&,A

*+,&40/56&A/)@9(2*$%&,2(&A/,9&*"(&E)8R(/&H&.,F

*"/,#@"& *,& *"(& (21& ,A& 40/56& ;/($01#($& 6IJ-K=L?

)21& ,2(& V#$*& B-*(/902)8& ,A& *"(& E)8R(/& H& .,F

*"/,#@"&*,&*"(&(21&,A&40/56&;/($01#($&66I-K=L?C

!,&7/,9,*(&#20A,/90*'&02&1(*(3*02@&40/56

1(8(*0,2& 1(/0P)*0P($%& )2& GH& *)@& +)$& )11(1& *,

3,2$*/#3*$&,A& *"(&40/56&/(@0,2$&38,2(1&02*,&)&<"#-

021#30.8(& 78)$901&+0*"& )2&70#28-9)*#"3+($7(30A03

/0.,$,9(-.02102@&$0*(C&O0@#/(&6&$",+$&$*()1'-$*)*(

8(P(8$&,A&$'2*"($0$&,A&GH-*)@@(1&A#88&8(2@*"&40/56

)21& *"(& *+,& B-*(/902)8& 40/56& 3,2$*/#3*$C& S2

*#9,/0@(2($0$& )$$)'$%& 40/56& )21& 40/56GH

(W#)88'&3,978(9(2*&)&<"#$%&1(8(*0,2&+"(/()$& *"(

B-*(/902)8& A/)@9(2*$& 1,& 2,*& ;1)*)& 2,*& $",+2?C

!)@@(1&P(/$0,2$&,A& A#88-8(2@*"&40/56&;/($01#($&<-

K=L?%& )21& *"(& *+,& B-*(/902)8& 40/56& A/)@9(2*$%

/($01#($& 6IJ-K=L& )21& /($01#($& 66I-K=L%& +(/(

02*/,1#3(1& 02& )#-,1& *,& )2&70#28-9)*#"3+ & BX=

1(/0P)*0P(& 3,2*)0202@& )& 78)$901& +0*"& !-TMH

.,/1(/$& A8)2R02@& *"(& 3,102@& $(W#(23($& ,A& $,8#.8(

@/((2& A8#,/($3(2*&7/,*(02& ;YOZ?&#21(/& *"(&3,2*/,8

,A&)&IX:&7/,9,*(/C

H& [+081-*'7(\& 7"(2,*'7(& $",+$& )& "0@"

8(P(8& ,A& */)2$0(2*& YOZ& (F7/($$0,2& 02& ?"92)"-,-

8*,)5-+"-,-& 8()P($%&+"(/()$& )& /(1#3*0,2& ,A&YOZ

(F7/($$0,2& /($#8*$& A/,9& )& A)08#/(& 02& !-TMH

*/)2$A(/C& Z/,*(02& 1,9)02$& *")*& )/(& $#AA030(2*& A,/

02*(/)3*0,2& .#*& 3)22,*& 3,978(9(2*& *"(&+081-*'7(

7/,*(02& 80R(8'& 10$/#7*& 40/5]!6::& */)2$7,/*(/

A#23*0,2& .'& .02102@& *,& $,9(& .#*& 2,*& )88& ,A& *"(

3,97,2(2*$&2(3($$)/'&A,/&)2&02*)3*& */)2$7,/*(/C&S2

(AA(3*%&*"(&2,2-7/,1#3*0P(&7/,*(02&A/)@9(2*$&*0*/)*(

,#*&7/,1#3*0P(&3,97,2(2*$%& 8()102@&*,&023,978(*(

)21& #2$*).8(& */)2$7,/*(/$C&H& /(1#3*0,2& ,A& ",& <"<2

A#23*0,2& 1#(& *,& 2,2-7/,1#3*0P(& 7"'$03)8

02*(/)3*0,2$& .(*+((2& */)2$7,/*(/& 3,97,2(2*$%

+,#81&3,2A0/9&,#/&*+,-"'./01&/($#8*$&*")*&02103)*(

*"(&B-*(/902)8&1,9)02&,A&40/56&0$&)&R('&/(@0,2&A,/

7/,*(02-7/,*(02& 02*(/)3*0,2$C& U)3"& 3,2$*/#3*& +)$

)$$($$(1&A,/&A#23*0,2&)*&8()$*&<>&*09($&02&I&$(7)/)*(

(F7(/09(2*$%&)21&/(7/($(2*)*0P(&1)*)&)/(&$",+2&02

O0@#/(&XC

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 7: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

!

!"##$#%&'()* +,-./* %01-%22%3* ,&* !"#$%* (4

()%* 5,#3$(61%* &'"(* 41%-4&* )73* 7#842(* 72* ),')* 7

#%9%#* 4:* ;!<* %01-%22,4&* 72* ()%* =4&(-4#* 2(-7,&2

>!,'"-%* ?@* 9%-2"2* AB* 7&3* CDE* F),2* -%1-43"=,G#%

3,::%-%&=%* G%(5%%&*+,-./*8%-43,1#4,3* 7&3*5,#3$

(61%*+,-./*#%9%#2*876*G%*3"%*(4*()%*2%&2,(,9,(6*4:

()%*262(%8*(4*()%*=4--%=(*2(4,=),48%(-6*4:*7##*F/HH

=4814&%&(2E*I4(%B*5%*&%9%-*4G2%-9%3* %01-%22,4&

4:*;!<*,&*&'"),&3"=%3**+",-#.!/"'01J*(),2*=4&(-4#

,2* 7#24* %9,3%&=%3* G6* ()%* :7=(* ()7(* =4&2(-"=(2* ()7(

14,24&* ()%* (-7&214-(%-* 34* &4(* -%2"#(* ,&* ),')*;!<

%01-%22,4&* '$) 23#$!#* >2%%* G%#45DE* !"-()%-84-%B

#45%-%3*F$AI@*(-7&2:%-*572*&4(*3"%*(4*7&*49%-7##

3%=-%72%*,&*F/HH*=4814&%&(*1-4(%,&*%01-%22,4&*4-

2(7G,#,(6* 72* 84&,(4-%3* G6* "&=)7&'%3* #%9%#2* 4:

+,-.K*,&*8%-43,1#4,3*2(-7,&2*G6*L%2(%-&*.#4((,&'

>37(7*&4(*2)45&DE

C01-%22,4&* 4:* +,-./* -%2,3"%2* /MN$OPQB

=4&(7,&,&'* ()%* L7#R%-* @* G40* -%3"=%3* ;!<

(-7&2:%-B*7&3*%01-%22,4&*4:*()%*+,-./*-%2,3"%2*//M$

OPQ* :-7'8%&(* 5,()4"(* ()%*L7#R%-* @* G40* '-%7(#6

-%3"=%3* ;!<* (-7&2:%-* >!,'"-%* ?.* 7&3* SDE* F)%2%

#7((%-* (54* -%2"#(2* 2"''%2(* ()7(* ()%* ,&(%-7=(,4&* 4:

()%2%* (54*+,-./* :-7'8%&(2*5,()*5,#3$(61%* F/HH

=4814&%&(2*#%732*(4*14,24&,&'*4:*F$AI@*(-7&2:%-E

F)%* 348,&7&(* &%'7(,9%* %::%=(* 4:* ()%* =4&2(-"=(

#7=R,&'* ()%*L7#R%-*@*G40*>!,'"-%*?SD* ,2*2(-4&'%-

()7&*()7(*4:*()%*=4&2(-"=(*=4&(7,&,&'*()%*L7#R%-*@

G40* >!,'"-%*?.DE*F),2* -%2"#(* ,2* %21%=,7##6*&4(7G#%

',9%&* ()7(* ()%*%01-%22,4&*4:* ()%*=4&2(-"=(*5,()4"(

()%* L7#R%-* @* G40* >!,'"-%* /B* #7&%2* ?* 7&3* TD* ,2

#45%-*()7&*()7(*4:*()%*=4&2(-"=(*5,()*()%*L7#R%-*@

G40* >!,'"-%* /B* #7&%2* M* 7&3* /DE* F)%* 2(-4&'%-

14,24&,&'*G6*()%*L7#R%-*@$8,&"2*=4&2(-"=(*87R%2

2%&2%* 72* (),2* =4&2(-"=(* ,2* #,R%#6* (4* G%* #%22* 7=(,9%B

7&3* ()"2* %0),G,(* 2(-4&'%-* ,&(%-:%-%&=%B* ()7&* ()%

=4&2(-"=(*()7(*2(,##*=4&(7,&2*()%*L7#R%-*@*G40E

AUHSVHHUWI

F)%*-%2"#(2*1-%2%&(%3*733-%22* ()%* #4=7(,4&

7&3* (414#4'6* 4:* ()%* S$(%-8,&"2* 4:* +,-./* ,&* 7

:-78%54-R* 4:* 2%9%-7#* 4()%-**+",-#.!/"'01*F/HH

=4814&%&(2E* W"-* :,&3,&'2* 2"114-(* 7* F/HH

2(-"=("-%*5)%-%,&*()%*)%078%-,=*+,-./*S$(%-8,&7#

3487,&*-%2,3%2*,&*()%*1%-,1#728*7&3*,&(%-7=(2*5,()

()%* +,-.KK* )%078%-* %8G%33%3* ,&* ()%* ,&&%-

8%8G-7&%E* F),2* =4&=#"2,4&* 2"1%-=%3%2* 4"-

1-%9,4"2* F/HH* 843%#* >PDB* 5),=)* 1-%3,=(%3* 7

=6(41#728,=*142,(,4&*:4-*+,-./*G72%3*24#%#6*4&*,(2

)484#4'6*(4*()%*(61%*U+*=4"1#,&'*1-4(%,&*+,-A/E

+,-./* (414#4'6* G72%3* 4&* 2%X"%&=%

7&7#62%2* 1-%3,=(2* :-48* Y%-4* >PD* (4* :4"-

(-7&28%8G-7&%* 3487,&2* >MPDB* 5),#%* 7#R7#,&%

1)421)7(72%* 2("3,%2* -%14-(* %,()%-* &4* F&$24,*

,&2%-(,4&2* ,&* +,-./* >MKBMND* 4-* 1%-,1#728,=

%0142"-%* 4:* ()%* -%',4&* 7-4"&3* -%2,3"%* /?Z* >K?DE

W"-* +,-./* (54$)6G-,3* ,&(%-7=(,4&2* 5,()* ()%

1%-,1#728,=* 3487,&2* 4:* +,-.P* 7&3* +,-.KZB* 7&3

4"-* :-7=(,4&7(,4&* 7&3*L%2(%-&* G#4(* 37(7* 2(-4&'#6

,&3,=7(%* 1%-,1#728,=* %0142"-%* 4:* ()%* +,-./* S$

(%-8,&7#*)%078%-,=*3487,&E*F),2*=4--%214&32*5,()

()%*7G49%*37(7*4&* ()%*7#R7#,&%*1)421)7(72%*:"2,4&

7(*-%2,3"%*/?ZE*U&*733,(,4&B*2("3,%2*5,()*3%(%-'%&($

24#"G,#,Y%3*("0./33#) %0'%*F/HH2*3%84&2(-7(%* ()7(

()%* +,-.P* 1%-,1#728,=* 3487,&* 7&3* 7* +,-./

)%078%-*%0,2(*,&*()%*278%*=481#%0*>QDE*W"-*+,-./

S$(%-8,&7#* 1%-,1#728,=* 1#7=%8%&(* ,2* :"-()%-

2"114-(%3* G6* 3%'-737(,4&* 4:* +,-./* ,&

*+",-#. ! / " ' 01 * 21)%-41#72(2* (-%7(%3* 5,()

1-4(%,&72%*[*>K/BK?DE*!,&7##6B*7&* ,&3%1%&3%&(*7&3

%#%'7&(* 72276B* F-U<B* 5),=)* 3%:,&%2* ()%* 4-3%-* 4:

,&(%-7=(,4&*G%(5%%&* ()%*F$2(-7&3* 7&3* =4814&%&(2

4:* ()%* F/HHB* 2"''%2(2* ()7(* ()%* F$2(-7&3* :,-2(

,&(%-7=(2*5,()*+,-A/*7&3*+,-.KK*,&*()%*=6(41#728

>K/DJ* ()%2%* 7"()4-2* )79%* 733,(,4&7#* '%&%(,=* 37(7

()7(*1#7=%* ()%* -%X",-%8%&(* :4-*+,-./*345&2(-%78

4:* ()%2%* ,&,(,7#* =4&(7=(2E* F)%2%* #7((%-* 37(7* 7-%

=4&2,2(%&(*5,()* 4"-* 722,'&8%&(* 4:* ()%*S$(%-8,&7#

@F<72%*3487,&*4:*+,-./*(4*()%*1%-,1#728E

F)%* 1-%2%&(* 2("3,%2* 34* &4(* 733-%22* ()%

(414#4'6*4:* ()%*I$(%-8,&"2*4:*+,-./E*C7-#6*54-R

2"''%2(%3* 7* 1422,G#%* 1%-,1#728,=* #4=7(,4&* 4:* 7

287##*I$(%-8,&7#*-%',4&*4:*+,-./*>-%2,3"%2*OT$P/D

1-%348,&7&(#6* G72%3* 4&* :,&3,&'2* ,&*56) .,3'* >K?DJ

)45%9%-B* ()%* 278%* 7"()4-2* 3%84&2(-7(%* ()7(* 7

-%',4&* 7-4"&3* 78,&4* 7=,3* NMO* ,2* =6(41#728,=* ,&

*+",-#. ! / " ' 01 E* L%* :794-* 7&* I$(%-8,&7#

=6(41#728,=* 722,'&8%&(* ,&* 4"-* 843%#* 3%2=-,G%3

G%#45* >!,'"-%* TDE* <4(%&(,7##6* 1-%3,=(,4&2* 4:

8%8G-7&%* (414#4'6* G72%3* 4&* :"2,4&2* (4* 4()%-

1-4(%,&2*876*&4(*7#5762*-%1-%2%&(*()%*&7(,9%*2(7(%E

F),2* =7"(,4&* ,2* %21%=,7##6* -%#%97&(* (4* 7* #7-'%

1-4(%,&* #,R%*+,-./*>OPQ* -%2,3"%2DB*5),=)*876*&4(

:4#3*1-41%-#6*5)%&* :"2%3* (4* 7#R7#,&%*1)421)7(72%

4 - * ! $'7#7=(42,372%E* F)"2B* G,4=)%8,=7#

:-7=(,4&7(,4&2*4:*&7(,9%*1-4(%,&2*3%2=-,G%3*)%-%,&B

72* 5%##* 72* G6* 4()%-* '-4"12* >QBKMBMOD* 876*84-%

7=="-7(%#6*-%:#%=(*()%,-*&7(,9%*#4=7#,Y7(,4&E

L%*4::%-* 4&%* :"-()%-* 14,&(* 4:* 3,2="22,4&E

<-4(%,&2* 876* 7--,9%* ,&* ()%* 1%-,1#728* G6* ()-%%

8%=)7&,282E*F)%*:,-2(* (54B*7*8%8G-7&%$217&&,&'

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 8: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

!

!"!#"$% !#&!% &'(#)$*% )'"% )$% +)$"% ,)+&-'*% ).% !#"

/$)!"-'% -'% !#"% /"$-/0&*+% )$% (0"&1&2"% ).% &'% 34

!"$+-'&0% *-2'&0% /"/!-,"5% &$"% 6"00% "*!&70-*#",8

9)6"1"$5% !#"$"% -*% &% !#-$,% /)**-7-0-!:8% ;% /$)!"-'

+&:% &$$-1"5% )$% )$-"'!% -!*"0.5% -'% !#"% /"$-/0&*+% 7:

+"&'*% ).% -'!"$&(!-)'% 6-!#% "-!#"$% +"+7$&'"4

&'(#)$",% )$% /"$-/0&*+-(% /$)!"-'*8% <#=*5% !#"% >4

!"$+-'=*%).%?-$@A%+&:%&$$-1"%-'%!#"%/"$-/0&*+%7:

-'!"$&(!-)'% 6-!#% ?-$@BB5% &*% 6"00% &*% ?-$@C% &',

?-$@BD8

<#"%/"$-/0&*+-(% 0)(&0-E&!-)'%).% !#"%?-$@A

;<F47-',-'2% ,)+&-'% /)*"*% &% /)!"'!-&0% G="*!-)'

.)$% #)6% !#-*% ,)+&-'% 2&-'*% &(("**% !)% ;<F8% H"

/$)/)*"% !6)% *("'&$-)*8% I'% !#"% .-$*!5% !#"% ;<F&*"4

,"/"',"'!% .='(!-)'%&',J)$% ()'.)$+&!-)'&0% (#&'2"

)((=$*% -'% !#"%(:!)/0&*+5%/$-)$% !)%&$$-1&0%).% !#"%>4

!"$+-'&0% /)$!-)'% ).% ?-$@A% &!% -!*% /"$-/0&*+-(

,"*!-'&!-)'8%I'%!#"%*"()',5%?-$@A%-*%&(!-1"%*)0"0:%-'

!#"% /"$-/0&*+-(% */&("5% &',% ;<F% -*% !$&'*."$$",

!#$)=2#% !#"% ("'!$&0% (#&''"0% ).% !#"% ?-$@BB

#"K&+"$-(% !$&'*+"+7$&'"% ()+/0"K% !)% !#"

/"$-/0&*+-(% #"K&+"$-(% ?-$@A% >4!"$+-'=*5

"+7",,",% 6-!#-'% !#"% !-2#!0:% L'-!% <AMM% ()+/0"K

NO-2=$"%PQ8% I'% !#-*% *("'&$-)5%;<F%6)=0,%')!%'"",

!)%($)**%&%+"+7$&'"%/"$%*"8%;*%!#"%!"#$%&'()#*+,

<AMM% !$&'*/)$!*% R3;5% -!% +&:% &0*)% !$&'*/)$!

'=(0")!-,"*%6-!#% *-+-0&$%7-)/#:*-(&0%/$)/"$!-"*% !)

'=(0"-(% &(-,% /)0:+"$*8% O)$% "K&+/0"5% !#"% ?-$@BB

#"K&+"$-(% ;<F&*"% 0-L"0:% -'!"$&(!*% 6-!#% ;<F

,=$-'2% !$&'*."$% ).% !#"% <4*!$&',% R3;% !#$)=2#% -!*

/)$"% NBAQ8%<#=*5%6"% .&1)$% &%+),"0%6#"$"%;<F% -*

&0*)% !$&'*."$$",% !#$)=2#% !#-*% /)$"5% ,-$"(!0:% !)

?-$@A5% !#"$"7:% "'&70-'2% ?-$@A% &(!-1-!:% -'% !#"

/"$-/0&*+8

?-$@BB% &**)(-&!"*% !-2#!0:% 6-!#% !#"% -''"$

+"+7$&'"S% TDU% ).% ?-$@BB% .$&(!-)'&!"*% 6-!#% !#"

-''"$%+"+7$&'"% &',% VDU% ).%?-$@BB% -*% .)=',% -'

!#"% (:!)/0&*+% NVWQ8% F$)!"-'&*"% X% *=*("/!-7-0-!:

*!=,-"*%,"+)'*!$&!"%!#&!%?-$@BB%-*%')!%"K/)*",%!)

!#"% /"$-/0&*+% NBYQ8% Z=$% !6)4#:7$-,% ,&!&

*=/"$-+/)*",% )'!)% )=$% ?-$@A% &',% ?-$@BB

*!$=(!=$&0% +),"0*% *#)6% )'"% .&("% ).% !#"% ?-$@A

#"K&+"$-(% ,)+&-'% -'!"$&(!*% 6-!#% !#"% 34!"$+-'&0

.&("%N$"*-,="*%B4BDPQ%).%!#"%?-$@BB%#"K&+"$8%<#-*

-'!"$&(!-)'% 6)=0,% !#"'% [()1"$\% &':% /)!"'!-&0

/"$-/0&*+-(%"K/)*=$"%).%?-$@BB5%&',%+&:%"K/0&-'

6#:%?-$@BB%-*%-++='"%!)%/$)!"&*"%,-2"*!-)'8%<#"

34!"$+-'&0% BDP% $"*-,="*% ).% 9FDY]Y5% !#"

-).*'$%&'()#/ 01.$#*% !"+/0&!"% .)$% )=$% ?-$@BB

#)+)0)2:% +),"05% ()4($:*!&00-E",% 6-!#% &

/)0:"!#:0"'"%20:()0%')'&+"$5%6#-(#%-',-(&!"*%!#&!

!#"% 34!"$+-'&0% $-'2% ).% 9FDY]Y% N&',% 0-L"0:

?-$@BBQ% -*% "+7",,",% -'% !#"% /#)*/#)0-/-,% 7-0&:"$

).% !#"% -''"$% +"+7$&'"% NBDQ8% ?-$@A4?-$@BB

()+/0"K% .)$+&!-)'% -*% ()'.-$+",% &0*)% 7:% ()4

-++=')%/$"(-/-!&!-)'%NBVQ8%<#"%!)/)0)2:5%/$)!"&*"

*=*("/!-7-0-!:5% ($:*!&00-E&!-)'5% .$&(!-)'&!-)'5% &',

!6)4#:7$-,% ,&!&% *=//)$!% )=$% /$)/)*&0% !#&!% !#"%>4

!"$+-'&05%/"$-/0&*+-(%?-$@A%#"K&+"$%*!&(L*%)'%!)/

).% !#"% -''"$4+"+7$&'"% "+7",,",% ?-$@BB

#"K&+"$8%@&$$"04.)$+-'2%*!&(L",%;<F&*"*%"K-*!%-'

)!#"$%7-)0)2-(&0%*:*!"+*5%-'(0=,-'2%!#"%>0/FJ>0/;

&',%>0/FJ>0/^%N/$)!"&*"J;<F&*"%()+/0"K"*Q%NADQ

&',%!#"%/WT%;;;%;<F&*"%&',%-!*%#)+)0)2*%NABQ8

?-$@A% "K-*!*% &*% &% #-2#0:% ()'*"$1",

()+/)'"'!% -'% <AMM*% NVCQ5% -',-(&!-'2% -!*

"1)0=!-)'&$:%&',%.='(!-)'&0% -+/)$!&'("8%R"0"!-)'*

).%?-$@A%&7)0-*#%7)!#%<4R3;% !=+)$% &',%<4/-0=*

.)$+&!-)'% NA]5AVQ5% 6#-0"% *)+"% ?-$@A%H&0L"$% ;

7)K%+=!&!-)'*% 0"&,% !)% 0)**%).%<4R3;%!$&'*."$%7=!

+&-'!"'&'("% ).% <4/-0=*% .)$+&!-)'% NWQ5% -',-(&!-'2

!#&!% ?-$@A% ()'!$-7=!"*% ,-.."$"'!-&00:% !)% /-0=*

.)$+&!-)'%&',%R3;%!$&'*."$8

I!% -*% *!$-L-'2% !#&!% &% 0&$2"% '=+7"$% ).

/#:*-(&0%-'!"$&(!-)'*%"K-*!%7"!6""'%!#"%>4!"$+-'=*

).% ?-$@A% &',% .)=$% ()+/)'"'!*% ).% !#"% <AMM5

?-$@B5% ?-$@C5% ?-$@BD% &',% ?-$@BB8% <#"*"% ,&!&

-+/0:% &% #-2#0:% ,:'&+-(% .='(!-)'&0% &',J)$% &'

-'!"2$&0% *!$=(!=$&0% $)0"% ).% ?-$@A8% <#"% +),"0% -'

O-2=$"%P%-00=*!$&!"*%#)6%!#"%?-$@A%>4!"$+-'=*%+&:

-'!"$&(!% 6-!#% !#"% /"$-/0&*+-(% ,)+&-'*% ).% ?-$@C

&',%?-$@BD5% !#"% -''"$4+"+7$&'"% .&("%).%?-$@BB5

&',% (:!)/0&*+-(% ?-$RA8% Z=$% +),"0% -'()$/)$&!"*

/$"1-)=*0:% ,"+)'*!$&!",% ?-$@C4?-$@BD5% ?-$@C4

?-$@BB5% &',% ?-$@BD4?-$@BB% :"&*!% !6)4#:7$-,

-'!"$&(!-)'*% NBT5AAQ5% &',% &% ?-$RA4?-$@BD

- ' ! " $ & ( ! - ) ' % , " ! " $+ - ' " , % 7 : % ( ) 4

-++=')/$"(-/-&!&!-)'% -'%!"#$%&'()#*+,%NBVQ8%H"

#:/)!#"*-E"%!#&!%!#"%>4!"$+-'=*%).%?-$@A%-'!"$&(!*

6-!#%?-$@C%&',%?-$@BD%-'%!#"%/"$-/0&*+%6#-0"%!#"

34!"$+-'=*% ).% ?-$@A% -'!"$&(!*% 6-!#% ?-$RA8% I'

*=//)$!5% ?-$RA5% ?-$@A% &',% ?-$@BB% &$"% ()4

-++=')/$"(-/-!&!",% NBVQ% &',% !#"% 34!"$+-'=*% ).

?-$@A% -'!"$&(!*% 6-!#% -!*"0.% NTQ% -'% !#"% 7&(!"$-&0

(:!)/0&*+8

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 9: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

!

!"#"!"$%"&

'( )*+,-./0(/1(/234456/!"##$%&'($)'*#+,'+-/!./37859

3( $:;:<./=(./:,>/!?@./%(/!(/234456/!.--$)'*#+,'+-/"./5A585B5

5( %C+<-D<E./F(/G(./:,>/%:-H:IE-./"(/2344J6/)+-$)./,#$0'+-/##./J'8K'

9( %C+<-D<E./F(/G(./LDM:N*+<./O(./O+<-C,:M??DC@./P(./G:N*Q?R-N<./&(./:,>/%:-H:IE-./"(/2344J6/1(("

2.3$)'*#+,'+-/"$./9J'89BJ

J( &HC+?>E+./S(./:,>/1:,N:./"(/2344J6/4-56/'7/"%./'83J

K( ):DEM:,./L(./%?<,./1(./0*+Q<,./!(./#<,,./!(/0(./=?II<HC./P(./S+<TT<DC-8G?,E-./&(./OC:,,:./L(.

U:+-C:II./U(./U?V?,./&(./&?,,C:MME+./"(/1(./&D*>C?IME./0(/G(./WE:D-./%(./:,>/">>@./&(/!(

234496/8"*-.'*$1*'76$2.6/&#./0'5B8'9'

A( 0:,;./X(/L(./YC?*./Z(/!(./S+:T./)(./:,>/%C+<-D<E./F(/G(/2'7776/)+-$)'*#+,'+-/&#./'3578'3J5

B( U<>>IED?,./!(./&[?I:,>E+./O(./#?IE@./G(./:,>/Y:MQ+@-N<./F(/2344J6/4#+*$859-$1*57$:*'$;$:$1/'(#.

'KBJ8'K74

7( W*:,./\(./%:+IE./L(./S:?./%(./&<]:,E-:,./0(./LI@./O(/L(./=?^^,E+./%(./O+:II./1(./0?MNE./$(./:,>

):+?,./%(/2344J6/<$0'+-$!=.//#)(./3K59783K5J7

'4( WE?./=(/G(./&:]]<>E-./&(/$(./=E++./L(/)(./1:,N:./"(./:,>/_:N-M:,./S(/234446/)+-$!.--/!./'9K'8

'9A3(

''( S?M<-8!*DC./#(/Z(./U?,H:I<:,./S(./FE+E`81*a*E./!(./S?,`:IE`./L(./%:QE`?,./"(./>E/I:/%+*`./#(.

:,>/%?II./U(/2344'6/859"#./%($./K5A8K9'

'3( b@E+./1(/U(./1E<^E./0(/0(./O??,<,./"(/P(./:,>/L+:]<,>./1(/234496/<$:9#"*9$0'+-/'%!./''85'

'5( LDM:N*+<./O(./%:-H:IE-./"(./:,>/%C+<-D<E./F(G(/234496/)+-$)'*#+,'+-/"%./''778'3''

'9( %:-H:IE-./"(./:,>/%C+<-D<E./F(/G(/234496/:*'.(*./&(%./''A48''A5

'J( 0:,;./X(/L(./:,>/%C+<-D<E./F(/G(/2'77A6/<$05*9.#'+-/'*$./9J589K3

'K( 0*+TEE./X(./0+:^E+./c(./Y*^:,./G(./%?,NI<,./0(/&(./:,>/Y:MQ+@-N<./F(/%(/2'7776/>.569/'"./'AK'8

'AKB(

'A( _:+>./0(/P(./0+:^E+./c(./Y*^:,./G(/!(./:,>/Y:MQ+@-N<./F(/%(/234436/4#+*$859-$1*57$:*'$;$:$1/$$.

''9758''J44

'B( 1I?-:./U(./Y*^:,./G(./):+?,./%(./:,>/Y:MQ+@-N<./F(/234446/<$05*9.#'+-/')#./595A8599J(

'7( X:<./X(/=(./0:CIQEHN./0(./%I:+N./"(/X(./S:[<R:I:./F(./F:-<?,./!(./_C:IE,./U(/%(./&D:II./!(/"(./:,>

&D:-N:R<H`./)(/G(/2'7776/4#+*$859-$1*57$:*'$;$:$1/$!./'9'J58'9'JB

34( 1:EMMI<./d(/O(/2'7A46/859"#./##*./KB48KBJ

3'( &HC:;;E+./=(./:,>/]?,/G:;?R./S(/2'7BA6/1(5-$0'+*=.//'!!./5KB85A7

33( 0E/U::;>./!(/L(./:,>/1*;DE,QE+;./)(/2'7BK6/<?$05*9.#'+-?/'!*./'4B58'4BJ

35( O*I>:*./S(/L(/2'77'6/<,/4-5(9$5(7$)'*#+,'5-$0'+-+@A./d,<]E+-<D@/?T/%:I<T?+,<:./)E+NEIE@.

)E+NEIE@./%L

39( &:MQ+??N./G(./:,>/!*--EII./0(/_(/2344'6/)+-.*"-5#$*-+('(@$B$5$-5,+#59+#A$/5("5-./5+>/">(/5

]?I-(./%?I>/&^+<,;/=:+Q?+/1:Q?+:D?+@/F+E--./%?I>/&^+<,;/=:+Q?+./$(W(

3J( &:I<./L(./:,>/)I*,>EII./X(/1(/2'7756/<$)+-$0'+-/#&%./AA78B'J

3K( S*EV./$(./:,>/FE<D-HC./U(/%(/2'77A6/C-.*9#+&=+#.6'6/')./3A'983A35

3A( G?,E-./X(/L(./Y?*./G(/W(./%?R:,./&(/_(./:,>/O[EI>;::+>(/2'77'6/1*95$!#A695--+@#$1/%*+,+-.+#/.

''48''7

3B( FEDDE+-E,./"(/#(./S?>>:+>./X(/0(./=*:,;./%(/%(./%?*HC./S(/&(./S+EE,QI:DD./0(/U(./UE,;./"(/%(./:,>

#E++<,./X(/"(/234496/<?$!+/&"9?$!=./?/#"./'K4J8'K'3

37( U*-CE;<:,./L(/!(./#*II,E+./O(/G(./O??,<,./"(/P(./:,>/$E-DE+./"(/_(/2'77K6/4#+*$859-$1*57$:*'$;$:

1/$&./A53'8A53K

54( Y:C+I./0(./_:;,E+./U(./)<-HC?T./O(./):@E+./U(./Y:]EH`./)(./)E+:,EN./L(./!*HNE,-D*CI./%(./Y:+TEI.

S(/"(./:,>/O?+:<M:,,./S(/2344J6/)'*#+,'+-+@A/'"'./59JJ859KA

5'( )E<[E+-QE+;E,./L(./&M<DC./&(/G(./:,>/=??@N::-./F(/G(/2'7796/4-56/'7/&#./3'383'B

53( 0:-./L(./:,>/Z<E./W(/=(/2'77B6/)+-$)'*#+,'+-/#*./94J89'9

55( ):+?,./%(./1I?-:./U(./YC?*./&(./:,>/Y:MQ+@-N<./F(/%(/2'77A6/<$05*9.#'+-/'*$./'3458'3'4

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 10: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

!"

!"# $%&'()*+(&+,-.#-/#,-0+1-2034'5(67,-8#-9#-:;<<"=-!"#$%&'()*+-!"#,-;>;;?;>;>

!@# $*''01&),-A#,-B05C7((,-/#,-D&3*+,-9#,-A*&+0'1,-E#-F#,-B0'&+,-9#,-0+1-G06(30+,-E#-:HII@=-,(*%

-$&+".%$/"0%)"1"0".-!$%,-"@<J?"JI;

!J# B07C*5,-K#,-G0'1,-L#,-M711C*)&+,-/#,-E'&((30++,-N#-E#,-0+1-2034'5(67,-8#-9#-:HIIJ=-,(*%"-$&+

.%$/"0%)"1"0".-!$&,-H@OH?H@O>

!># 90(P0C*(,-Q#,-0+1-9%'7()7*,-8#-N#-:HII"=-,(*%"-$&+".%$/"0%)"1"0".-!$!,-;>HHO?;>H!!

!O# 90&,-$#-B#,-0+1-K07*',-M#-R#,-N'#-:HII;=-2)%(*3)*+*45-!'",-!HI;?!H;"

!<# /0(%6&S0,-K#,-KTU17P%,-E#-M#,-0+1-9%'7()7*,-8#-N#-:;<<>=-!"#$%&'()*+-!"(,-@O!?@<;

"I# E'74U+,-F#,-V734*',-M#-K#,-9%7+W,-/#,-KT'0+W*'(,-/#,-X7*47W,-V#-M#,-0+1-R&U'5,-G#-F#-:HII@=-!

#)*+"67'8-%)$,-;J;O@?;J;<J

";# Y0C*,-/#-L#-:HIII=-!"6'++"#)*+-!*$,-X;!?;<

"H# B*'W*',-B#-/#,-0+1-9%'7()7*,-8#-N#-:;<<!=-!"#$%&'()*+-!"*,-;>H!?;>!"

"!# K0WUC*+6&,-Q#,-K0WUC*+6&,-Y#,-9%*+,-N#,-0+1-9%'7()7*,-8#-N#-:HII;=-!"#$%&'()*+-!)&,-@O;!?@OH@

""# L0(,-F#,-0+1-Z7*,-.#-R#-:HIII=-!"#$%&'()*+-!)%,->@O?>J!

XDD$[D$QK

\-$%*(*-0U)%&'(-P&+)'74U)*1-*]U0CC5#

^-G*- )%0+6- )%*-9[/-B7&C&W7P0C- _30W7+W-X0P7C7)5- 0)-`9-B*'6*C*5- a&'-37P'&(P&T5-0((7()0+P*#-ML-b0(

(UTT&')*1- 45- 0- `+7S*'(7)5- &a- 90C7a&'+70- X*CC&b(%7T- 0+1- DL- 0+1- /M- b*'*- (UTT&')*1- 45- 0- [0)7&+0C

KP7*+P*-X&U+10)7&+-W'0+)-I!"!@JJ-)&-82#

H#- $%*- 044'*S70)7&+(- U(*1- 0'*-$"KK,- )5T*- _Y- (*P'*)7&+- (5()*3c-.Q8L,- 5*0()- *d)'0P)- T*T)&+*- 1*d)'&(*

3*17U3c- 8MKX,- T%*+5C3*)%5C(UCa&+5C- aCU&'71*c- QL$F,- *)%5C*+*17037+*)*)'00P*)7P#$%&'(-$BK$,-$'7(?

4Uaa*'*1- (0C7+*- $b**+?HIc- /BK,- '74&(&3*?47+17+W- (7)*c- RF- %*30WCU)7+7+c- 8YLX,- T&C5S7+5C71*+*

17aCU&'71*c- AB,- AU'70?B*')0+7- 3*17U3c- KLK?8FEQ,- (&17U3- 1&1*P5C- (UCa0)*- T&C50P'5C0371*- W*C

*C*P)'&T%&'*(7(c- MQK,- H?:+?3&'T%&C7+&=?*)%0+*(UCa&+7P- 0P71c- EX8,- W'**+- aCU&'*(P*+)- T'&)*7+c- FFF,

F$80(*(-F((&P70)*1-b7)%-17S*'(*-P*CCUC0'-FP)7S7)7*(c-$?L[F,-)'0+(a*''*1-L[Fc

X_E`/Q-AQEQ[LK

X7W#-;+-KU330'5-&a-)b&?%54'71-7+)*'0P)7&+(-4*)b**+-Y7'B"-0+1-&)%*'-Y7'B-T'&)*7+(#-.,-D+-)%*-'7W%)-(71*

&a- )%*- a7WU'*,- )%*- &'7W7+- &a- *0P%- 0''&b- 7+17P0)*(- )%*- 407),- 0+1- )%*- )7T- &a- )%*- 0''&b- )%*- T'*5- &a- )%*

7+)*'0P)7&+#-BC0P6-%&'7e&+)0C-C7+*(-'*T'*(*+)-)%*-037+&-0P71-(*]U*+P*,-b7)%-+U34*'(-1*30'P0)7+W-'*(71U*

+U34*'(#-/*W7&+(-&a- 7+)*'0P)7&+-4*)b**+-)b&-T'&)*7+(-0'*-(%&b+-45-(b0)%(-&a- C0S*+1*'-0+1-W'**+#-$%*

10'6*'-P&C&'-(b0)%(-&PPU'-b%*+-)b&-7+)*'0P)7&+(-&S*'C0Tc- )%*(*-'*aC*P)- )%*-37+730C-e&+*(-&a- )b&?%54'71

7+)*'0P)7&+#- #,- /*W7&+(- &a-Y7'B"- )%0)- 7+)*'0P)*1-b7)%- &)%*'-Y7'B- T'&)*7+(- 0'*-30TT*1- 4*C&b- 0- C7+*0'

'*T'*(*+)0)7&+-&a- aUCC?C*+W)%-Y7'B"#- _+-T0'*+)%*(*(-0'*- )%*-Y7'B"- '*(71U*(- )%0)- 7+)*'0P)-b7)%- )%*-+03*1

Y7'B-T'&)*7+c-a&'-*d03TC*f-Y7'B;-:"!H?@HH=-7+17P0)*(-)%0)-Y7'B"-:'*(71U*(-"!H?@HH=-7+)*'0P)(-b7)%-Y7'B;#

X7W#-H#-L&307+(-&a-7+)*'0P)7&+-30TT*1-&+)&-3&1*C(-&a-Y7'B"-0+1-Y7'B;;#-.,-M&1*C-&a-)%*-P'5()0CC7e*1-9:

;5+*()-R8I@H@-173*'-&+-)%*-C*a)-:7+-TU'TC*=-P&3T0'*1-)&-0-3&1*C-&a-)%*-.:"&<8'=$%)'>?-Y7'B;;-173*'-&+

)%*-'7W%)-:7+-4CU*=#-#,-M&1*C-&a-)%*-9:";5+*()-R8I@H@-%*d03*'-&+-)%*-C*a)-P&3T0'*1-)&-0-3&1*C-&a-)%*-.:

&<8'=$%)'>?-Y7'B;;-%*d03*'-&+-)%*-'7W%)#-6,-$&T-C*a)f-K71*-S7*b-&a-0-Y7'B"-9?)*'37+0C-%*d03*'-:0(-0

'744&+-170W'03=-b7)%-)%*-T'*5-'*W7&+-&a-7+)*'0P)7&+-7+-)*0C#-Y7'B"-T'&)'U1*(-7+)&-)%*-T*'7TC0(37P-(T0P*-:8=#

$&T- '7W%)f- B&))&3- S7*b- &a- 0- (T0P*?a7CC7+W- 3&1*C- &a- )%*- Y7'B"- %*d03*'- b7)%- )%*- '*W7&+- &a- Y7'B;;

7+)*'0P)7&+-7+-)*0C#-B&))&3-C*a)f-K71*-S7*b-&a-0-Y7'B;;-%*d03*'-:0(-0-'744&+-170W'03=-b7)%-)%*-407)-'*W7&+

&a-7+)*'0P)7&+-7+-4CU*#-Y7'B;;-T'&)'U1*(-7+)&-)%*-P5)&TC0(3-:9=#-B&))&3-'7W%)f-$&T-S7*b-&a-0-(T0P*?a7CC7+W

3&1*C-&a-)%*-Y7'B;;-%*d03*'-b7)%-)%*-'*W7&+-&a-Y7'B"-7+)*'0P)7&+-7+-4CU*#-$%*-4C0P6-(P0C*-40'(-'*T'*(*+)

;I-0+W()'&3(#

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 11: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

!!

!"#$% &$% !'()*"+,(*"+,% +-% -.//0/1,#*2% 3"'45% (,6% *21% 3"'45% 70*1'8",(/% -'(#81,*$% 91:*1',% ;/+*:% +-% !"

#$%&'()*&+,% )1//:% 1<='1::",#% *21% 218(#/.*",",% >?@A0*(##16% 70*1'8",.:% +-% 3"'45% >'1:"6.1:% 5&B0CDEA

)+,:*'.)*%",%*'(,:%*+%F"/60*G=1%3"'45$%H+/1)./('%8(::1:%>",%IJ(A%('1%*+%*21%/1-*%+-%1()2%;/+*$%!'()*"+,:%('1

",6")(*16%(;+K1%*21%;/+*:L%(:%F1//%(:%*21%='1:1,)1%>MA%+'%(;:1,)1%>0A%+-%*21%",6.)1'%()1*+:G'",#+,1%>@NA$%!L

4/+*%='+;16%F"*2%*21%3"'45%O0*1'8",(/%(,*";+6GL%F2")2%'1)+#,"P1:%*21%O0*1'8",.:%+-%3"'45%>'1:"6.1:%Q0

QR&A$%S21%(''+F21(6% ",6")(*1:% *21%-.//0/1,#*2%3"'45%='+*1",$%-L%4/+*%='+;16%F"*2% *21%3"'45%70*1'8",(/

(,*";+6GL%F2")2% '1)+#,"P1:% *21%70*1'8",.:% +-%3"'45% >'1:"6.1:% T&B0CDEA$%.L%4/+*%='+;16%F"*2% *21%?@

(,*";+6G$%/L%4/+*% ='+;16%F"*2%3"'4E% (,*";+6G$%@''+F21(6:% ",6")(*1% -.//0/1,#*2%3"'45% >!AL% -.//0/1,#*2

3"'45%=/.:%*21%70*1'8",.:%+-%3"'45%>-AL%*21%70*1'8",.:%+-%3"'45%>.AL%(,6%3"'4E%>/A$

!"#$%5$%U<='1::"+,%+-%3"'45%(,6%61'"K(*"K1:$%91:*1',%;/+*:%+-%?@0*(##16%3"'45%61'"K(*"K1:$%H+/1)./('

8(::1:%>",%VJ(A%('1%*+%*21%/1-*$%W(,1%,.8;1':%('1%(;+K1%*21%;/+*:$%W()I%+-%",6.)*"+,%>0A%+'%",6.)*"+,%>MA

F"*2%()1*+:G'",#+,1%":% ",6")(*16$%@//%-'(#81,*:%F1'1%)/+,16%",*+%(,%1<='1::"+,%=/(:8"6%(,6%=/()16%",%(

F"/60*G=1L% 7XDL% ;()I#'+.,6$% W(,1:% Q% (,6% BY% ?@03"'45% >'1:"6.1:% Q0CDEA$% W(,1:% &% (,6% 5Y% ?@03"'45

>'1:"6.1:% 5&BZCDEA$% W(,1:% X% (,6% TY%?@03"'45% >'1:"6.1:% 55&ZCDEA$% W(,1:% C% (,6% DY%?@% (/+,1$%7'+::0

'1()*"K1%='+*1",>:AL%K":";/1%",%;+*2%",6.)16%(,6%.,",6.)16%/(,1:L%'.,%(*%*21%;'+8+=21,+/%;/.1%6G1%-'+,*%",

(//%/(,1:$%!+'%/(,1:%C%(,6%DL%1/1)*'+=2+'1:":%F(:%-+'%(%:2+'*1'%*"81%*+%(//+F%61*1)*"+,%+-%*21%?@%*(#%F2")2

'.,:%-(:*1'%*2(,%*21%6G1%-'+,*$

!"#$%X$%J+8",(,*%,1#(*"K1%1--1)*:%+-%3"'45%81'+6"=/+"6:$%@//%3"'45%-'(#81,*:%F1'1%1<='1::16%",%(%F"/60

*G=1%*'(,:=+'*1'%;()I#'+.,6$%S21%2+'"P+,*(/%'1)*(,#/1:%'1='1:1,*%3"'45%'1:"6.1:[%9(/I1'%@%(,6%9(/I1'%4

;+<1:%('1%",%+'(,#1$%\8(#1:%+-%]!^%1<='1::"+,%",%0*)1#*(+(23&+#4(%*(+(2=/(,*:[%*21%6('I%)"')/1:%('1%*21

;()*1'"(/% :.:=1,:"+,% ",-"/*'(*"+,% 2+/1:$%!L% !.//% /1,#*2% 3"'45$% -L% 3"'45% >'1:"6.1:% 5&B0CDEA$%.L% 3"'45

>'1:"6.1:%55&0CDEA$%/L%?@%*(#%(/+,1$%5L%4()I#'+.,6%:*'(",$

!"#$%T$%H+61/%+-%3"'45% ",*1'()*"+,:%F"*2%3"'4QQL%3"'J5L%3"'4D%(,6%3"'4QR$%S21%21<(81'")%3"'45%70

*1'8",.:%",*1'()*:%F"*2%",,1'0818;'(,1%3"'4QQ%(,6%=1'"=/(:8")%3"'4D%(,6%3"'4QR$%3"'4D%(,6%3"'4QRL

",%*.',L% ",*1'()*%F"*2%1()2%+*21'%(,6%3"'4QQ$%3"'4QR%",*1'()*:%F"*2%3"'J5%>Q&A$%91%='+=+:1%O0*1'8",(/

3"'45%",*1'()*:%F"*2%)G*+=/(:8")%'1#"+,:%+-%3"'J5%(,6%3"'4QR$

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 12: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

!"

#!$

%#& '("

)*+,!-./&0

)*+,$-#(#0

)*+,!-./&0

)*+,$$-%!!0

)*+,/-#%.0

)*+,!-./&0

)*+,$"-%..0

!!% ('!

$"'

%#& '("!%# ( " '($

)*+,$-#(#0' $!'

%.

$

$&/

#(#

$$ $$

!"$

$((

!"$

$(

#&

#%.#%.'" #%' #%.

(#" (## '($

#(#

(##

/ #(#

'

'$

'!%#

'$ $

1

,$2-!%#3(##0

,$2-(#"3'($0

,/2-%#&3!"$0

,$"2-%#&3'("0

,$$2-!!%3('!0

45678+12,9:2-!%%3!!"0

45678+2,2,9:2-'$&3'%(0

,

Figure 1

!"

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 13: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

A

C

B

P

C

Figure 2

13

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 14: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

kDa – – – –+ +++

33

10072

55

4055

40

11481

ASTotalPerip

lasm

Cytoplasm

Membrane

A

B

C

D

20

Figure 3

!"

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 15: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

!"#$

%& ' ( ' ( ' ( ' () *+ $ ,- " !

*.#/-/#/0

+#)

Figure 4

!"

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 16: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

!

"

#

$

%

Figure 5

!"

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 17: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

!" !"

# #

!"

$%

%

!!

Figure 6

!"

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from

Page 18: 01!*-2,345',3'!)*' !#$%!&'(#)*+ '*+(,!&)(-. '+,-.67/'!&*!"#$%&'()$)* + , -. -/ 0 1 )2 34)56#%7$"%76)89:'()$&5 2 ; 9

Olga Draper, Rebecca Middleton, Michaeleen Doucleff and Patricia C. Zambryskisecretion system

Topology of the VirB4 C-terminus in the agrobacterium tumefaciens VirB/D4 type IV

published online October 12, 2006J. Biol. Chem. 

  10.1074/jbc.M606403200Access the most updated version of this article at doi:

 Alerts:

  When a correction for this article is posted• 

When this article is cited• 

to choose from all of JBC's e-mail alertsClick here

Supplemental material:

  http://www.jbc.org/content/suppl/2006/10/17/M606403200.DC1

by guest on January 26, 2020http://w

ww

.jbc.org/D

ownloaded from