1

细胞信号转导

2

cAMP 的第二信使作用 / 信息在细胞内的逐级放大

催化亚基

蛋白激酶 A（ 4聚体）

调节亚基 2聚体

＋

R

CC

R

RRC C cAMP

cAMPcAMP

cAMP

多种靶蛋白的某些丝 /苏氨酸残基磷酸化

第二信使 cAMP

ATP

腺苷酸环化酶

G 蛋白 s

受体 第一信使

3

磷酸化共价修饰代谢途径中的关键酶 糖原合成酶：抑制糖原合成 糖原磷酸化酶 b激酶 糖原磷酸化酶 a ：促进糖原分解

特点：一 被磷酸化共价修饰的蛋白发生生物效应的空间位

置位于胞浆。

二 调节速度相对快。

4

肾上腺素 /β 受体或胰高血糖素 / 受体

G 蛋白 G 蛋白

ACiACa

ATPcAMP

PKAiPKAa

磷酸化酶 b 激酶

磷酸化酶 b 激酶 -P磷酸化酶 a磷酸化酶 b

磷蛋白磷酸酶 -1PKAa

磷蛋白磷酸酶 -1

抑制物 -P

磷蛋白磷酸酶 -1PKAa

抑制物ATP Pi

+

_

_

_

葡萄糖 ATP

_ _

ATP

Pi

ATP Pi

AMP

糖原合酶

糖原合酶 -P

Pi ATP

磷蛋白磷酸酶 -1 PKAa_

糖原合成分解代谢途径相关酶磷酸化共价修饰调节的相互协调

G1P

糖原

Mg2+

无活性

无活性

无活性

无活性

5

cell membrane Gs/i

cAMP PKAc

P1

ACR

下丘脑的释放激素

ATP

TGACGTCA geneCREB P

CREB

RNA

肾上腺素

胰高血糖素垂体促激素 抗利尿激素 绒毛膜促性腺激素甲状旁腺素

大多数肽类激素

AMP

PDE+

_茶碱

胰岛素Ca2+

CaM

儿茶酚胺， α2 受体， Gi

儿茶酚胺， β 受体

生长抑素， α2 受体， Gi

cell in

cell out

TPK

R

信息物质 -X

，信息物质 -X

信息物质 -Y信息物质 -Y ？

生长因子癌蛋白

细胞因子

cAMP 介导的信息跨膜传递

生化效应：靶蛋白丝 /苏氨酸的磷酸化 抑制糖原合成，促进糖原分解 促进脂肪分解，抑制脂肪酸合成 去除对相关基因转录的阻遏，并同时加速转录与翻译过程 使心肌肌原蛋白易于结合 Ca2+ ，加速心肌肌浆网膜蛋白摄 Ca2+

改变微管蛋白的构象与功能，影响细胞分泌 影响细胞膜蛋白的构象，改变膜对水和离子的通透性

胰岛素， Gi

抗血管紧张素Ⅱ， Gi

促肾上腺皮质激素

Mg2+

ATP

6

7

8

9

10

11

Src Homology domain, SH1, SH2 and SH3

SH1: Tyrosine kinase domain

SH2: recognizes and binds with high affinity to peptide motifs containing a phosphotyrosine (pTyr) and involves Signal transduction.

SH3: binds to proline-rich ligands with moderate affinity and selectivity, preferentially to PxxP motifs; they play a role in the regulation of enzymes by intramolecular interactions, changing the subcellular localization of

12

For examples

SH3

SH3

SH3 SH3

SH3

SH2

SH2

SH2

SH2 SH2

SH2

SH1, tyrosine kinase

Phospholipase C Phospholipase C

GTPase activating protein

Src

Grb2

GAP

PLC 1

13

1 ME AIAKY*DF*KATADDE*LSS11FKRGDILKVLNEECDQN

36 W*YKAELNGKDGFIP1*1*KNY*IE MKP22H P3WFFGKIP4RAKA

71 EEMLSS2KQRHDGAFLIRESS3ESS4AP5GDFSS5LSS6VKFGNDV

106 QHFKVLRDGAGKYFLWVVKFNSS7LNELVDYHRSS8TSS9VSS10R143 NQQIFLR DIEQVP66QQP77T YVQALF*DF*DP88QEDGE*LGFRRG

181 DFIHVMDNS1111DP99NW*WKGACHGQTGMFP10*10*RNY*VTP1111 VNRNV

Primary structure of Grb2 protein

SH3

-3SH3

-2SH2

+7

3-54 59-149 160-212

+2

* proline-rich ligand binding site

14

ME A3IAKYDFKATADDELSFKRGDILKVLNEECDQNWYKAELNGKDGFIPKNYIE54 MKP57H P59WFFGKIPRAKAEEMLSKQRHDGAFLIRESESAPGDFSLSVKFGNDVQHFKVLRDGAGKYFLWVVKFNSLNELVDYHRSTSVSRNQQIFLR149 DIEQVP155QQP158T Y160VQALFDFDPQEDGELGFRRGDFIHVMDNSDPNWWKGACHGQTGMFPRNYVTP212 VNRNV

15

MKPHPWFFGKIPRAKAEEMLSKQRHDGAFLIRESESAPGDFSLSVKFGNDVQHFKVLRDGAGKYFLWVVKFNSLNELVDYHRSTSVSRNQQIFLRDIEQVP

Tertiary structure of SH2

16

Molecular Weight: 529.481 g/molMolecular Formula: C21H32N5O9PHydrogen Bond Donor Count: 7Hydrogen Bond Acceptor Count: 9Rotatable Bond Count: 14 , Charge: 0 Tautomer Count: 32, Heavy Atom Count: 36 Topological Polar Surface Area: 240XLogP: -3.3, Exact Mass: 529.194MonoIsotopic Mass: 529.194Complexity: 855, Isotope Atom Count: 0Defined Atom StereoCenter Count: 3Undefined Atom StereoCenter Count: 0Defined Bond StereoCenter Count: 0Undefined Bond StereoCenter Count: 0Covalently-Bonded Unit Count: 1

IUPAC Name: [4-[(2R)-2-[[(2S)-1-[[(2S)-1,4-diamino-1,4-dioxobutan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]carbamoyl]-4-methylamino-4-oxobutyl]phenyl] dihydrogen phosphateCanonical SMILES: CC(C)C(C(=O)NC(CC(=O)N)C(=O)N)NC(=O)C(CC1=CC=C(C=C1)OP(=O)(O)O)CC(=O)NCIsomeric SMILES: C(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N)NC(=O)[C@H](CC1=CC=C(C=C1)OP(=O)(O)O)CC(=O)NCInChI: InChI=1/C21H32N5O9P/c1-11(2)18(21(31)25-15(19(23)29)10-16(22)27)26-20(30)13(9-17(28)24-3)8-12-4-6-14(7-5-12)35-36(32,33)34/h4-7,11,13,15,18H,8-10H2,1-3H3,(H2,22,27)(H2,23,29)(H,24,28)(H,25,31)(H,26,30)(H2,32,33,34)/t13-,15+,18+/m1/s1/f/h24-26,32-33H,22-23H2

17

Molecular Weight: 46.0254 g/molMolecular Formula: CH2O2

Hydrogen Bond Donor Count: 1Hydrogen Bond Acceptor Count: 2Rotatable Bond Count: 0Topological Polar Surface Area: 37.3XLogP: -0.3Exact Mass: 46.0055MonoIsotopic Mass: 46.0055Heavy Atom Count: 3Charge: 0Complexity: 10.3Isotope Atom Count: 0Defined Atom StereoCenter Count: 0Undefined Atom StereoCenter Count: 0Defined Bond StereoCenter Count: 0Undefined Bond StereoCenter Count: 0Covalently-Bonded Unit Count: 1IUPAC Name: formic acidCanonical SMILES: C(=O)OInChI: InChI=1/CH2O2/c2-1-3/h1H,(H,2,3)/f/h2H

18

MKPHPWFFGKIPR67AKAEEMLSKQRHDGAFLIR86ESESAPGDFSLSVKFGNDVQH107F108K109VLRDGAGKYFLWVVKFNSLNELVDYH135RSTSVSRNQQIFLRDIEQVP

R67

R86

H107 K109

pTyr binding pocket

F108

H135

unique binding pocket

19

Grb2 binds glycerol

glycerol

20

green A I A K Y* D F* K A

red Y V Q A L F* D F* D P

green T A D D E* L SS F K R

red Q E D G E* L G F R R

green G D I L K V L N E E

red G D F I H V M D N S

green C D Q N W* Y K A E L

red D P N W* W K G A C

green N G K D G F I P** K N

red H G Q T G M F P** R N

Consensus or similar sequence

Entire: 55.56%, core: 83.33% green Y* I E

red Y* V T P

SH3 of Grb2

21

Grb2, N-Terminal Sh3 DomainComplexed With Sos-A Peptide (Nmr, 29 Structures)

Sos-A: SPLLPKLPPKTYKRE(Mouse )(Homo sapiens, Sos-A : SPHLPKLPPKTYKRE)

SH3: GSRRASVG SMEAIAKYDFKATA D DELSFKR G DILKVL NEECDQ NWYKAELNGKDFIPK N YIEMKP HPEFIVTD

22

Sos-A: SPLLPK6LPPK10TYKRE

SH3:GSRRASVGSMEAIAKYDFKATAD23D24E25*LSFKRGDILK

VLNEECD42QNWYKAELNGKDFIPKNYIEMKPHPEFIVTD

--- D23D24E25*

+ K6

+ K10

- D42

23

Sos-A: SP2L3L4P5KL7P8P9KTYKRE

SH3:GSRRASVGSMEAIAKY16*D17F18*K19A20TADDE*LS

FKRGDILKVLNEECDQNW*YKAELNGKDF55I56P57*KNY*IEMKPHPEFIVTD

P2L3L4P5

L7P8P9

Y16D17F18K19A20

F55I56P57

24

MEAIAKY*DF*KATADDE*LSFKR

GDILKVLNEECDQNW*YKAELNGK

DGFIP*KNY*IEMKPHPWFFGKIPR

AKAEEMLSKQRHDGAFLIRESESAP

GDFSLSVKFGNDVQHFKVLRDGAG

KYFLWVVKFNSLNELVDYHRSTSVS

RNQQIFLRDIEQVPQQPTYVQALF*

DF*DPQEDGE*LGFRRGDFIHVMD

NSDPNW*WKGACHGQTGMFP*RN

Y*VTPVNRNV

Proline-rich ligand binding site on SH3 of Grb2

25

MQQAPQPYEFFSEENSPKWRGLLVSALRKVQVQVHPTLSANEESLYYIEELIFQLLNKLCMAQPRTVQDVE

ERVQKTFPHPIDKWAIADAQSAIEKRKRRNPLLLPVDKIHPSLKEVLGYKVDYHVSLYIVAVLEYISAD

ILKLAGNYVFNIRHYEISQQDIKVSMCADKVLMDMFDQDDIGLVSLCEDEPCSSGELNYYDLVRTEIAEERQ

YLRELNMIIKVFREAFLSDRKLFKPSVYEKIFSNISDIHELTVKLLGLIEDTVEMTDESSPHPLAGSCFEDLA

EEQAFDPYETLSQDILSPEFHEHFNKLMARPAVALHFQSIADGFKEAVRYVLPRLMLVPVYHCWHYFELLK

QLKACSEEQEDRECLNQAITALMNHQGSMDRIYKQYSPRRRPGDPVCPFYSHQLRSKHLAIKKMNEIQK

NIDGWEGKDIGQCCNEFIMEGPLTRIGAKHERHIFLFDGLMISCKPNHGQTRLPGYTSAEYRLKEKFVMRKI

QICDKEDTCEHKHAFELVSKDENSIIFAAKSAEEKNNWMAALISLHYRSTLDRMLDSVLLKEENEQPLRLP

SPEVYRFVVKDSEENIVFEDNLQSRSGIPIIKGGTVVKLIERLTYHMYADPNFVRTFLTTYRSFCKPQELLSL

LIERFEIPEPEPTDADKLAIEKGEQPISADLKRFRKEYVQPVQLRVLNVFRHWVDHHYYDFERDLELLERL

ESFISSVRGKAMKKWVESIAKIIRRKKQAQANGVSHNITFESPPPPIEWHISKPGQFETFDLMTLDPIEIARQ

LTLLESDLYRKVQPSELVGSVWTKEDKEINSPNLLKMIRHTTNLTLWFEKCIVEAENFEERVAVLSRIIEILQ

VFQDLNNFNGVLEIVSAVNSVSVYRLDHTFEALQERKRKILDEAVELSQDHFKKYLVKLKSINPPCVPFFGI

YLTNILKTEEGNNDFLKRKGKDLINFSKRRKVAEITGEIQQYQNQPYCLRIEPDMRRFFENLNPMGSASEKE

FTDYLFNKSLEIEPRNCKQPPRFPRKSTFSLKSPGIRPNTGRHGSTSGTLRGHPTPLEREPCKISFSRIAET

ELESTVSAPTSPNTP63STPPVSASSDLSVFLDVDLNSSCGSNSIFAPVLLPHSKSFFSSCGSLHKLSEEPLIPP

PLPPRKKFDHDASNSKGNMKSDDDPPAIPPRQPPPPKVKPRVPVPTGAFDGPLHSPPPPPPRDPLP

DTPPPVPLRPPEHFINCPFNLQPPPLGHLHRDSDWLRDISTCPNSPSTPPSTPSPRVPRRCYVLSSSQNN

LAHPPAPPVPPRQNSSPHLPKLPPKTYKRELSHPPLYRLPLLENAETPQ

SOS2(son of sevenless homolog 2, Homo sapiens)Proline rate: total, 9.3%(124); front part 1088 aa, 5.7%; back part 244 aa, 25.4%

26

C

O

OCH

CHO

H

H

CH O C

OH

O

OH

HO

O

O

OH

P

P

P

O

O

O

OH

O-

HO

OH

O-

O-

Inosital triphosphate

Diacylglycerol

27

Solution Structure of The C2h2 Type Zinc-Binding Domain of Human Zinc Finger Protein 292 [mmdbId:35874]

B side

A ahead

(RKIRRHKKEKEEKK)gssgssgRKKPVSQSLEFPTRYSPYRPYR

CVHQGCFAAFTIQQNLILHYQAVHKSDLPAFSAEVEEESgpssg(AGKESEETETKQ)

C C

H H

Zn2+

H

H

C

C

44

12

28

MADEEAEQERLSCGEGGCVAELQRLGERLQELELQLRESRVPAVEAATDYCQQLCQTLLEYAEKWKTSEDPLPLLEVYTVAIQSYVKARPYLTSECENVALVLERLALSCVELLLCLPVELSDKQWEQFQTLVQVAHEKLMENGSCELHFLATLAQETGVWKNPVLCTILSQEPLDKDKVNEFLAFEGPILLDMRIKHLIKTNQLSQATALAKLCSDHPEIGIKGSFKQTYLVCLCTSSPNGKLIEEISEVDCKDALEMICNLESEGDEKSALVLCTAFLSRQLQQGDMYCAWELTLFWSKLQQRVEPSIQVYLERCRQLSLLTKTVYHIFFLIKVINSETEGAGLATCIELCVKALRLESTENTEVKISICKTISCLLPDDLEVKRACQLSEFLIEPTVDAYYAVEMLYNQPDQKYDEENLPIPNSLRCELLLVLKTQWPFDPEFWDWKTLKRQCLALMGEEASIVSSIDELNDSEVYEKVVDYQEESKETSMNGLSGGVGANSGLLKDIGDEKQKKREIKQLRERGFISARFRNWQAYMQYCVLCDKEFLGHRIVRHAQKHYKDGIYSCPICAKNFNSKETFVPHVTLHVKQSSKERLAAMKPLRRLGRPPKITTTNENQKTNTVAKQEQRPIKKNSLYSTDFIVFNDNDGSDDENDDKDKSYEPEVIPVQKPVPVNEFNCPVTFCKKGFKYFKNLIAHVKGHKDNEDAKRFLEMQSKKVICQYCRRHFVSVTHLNDHLQMHCGSKPYICIQMKCKAGFNSYAELLTHRKEHQVFRAKCMFPKCGRIFSEAYLLYDHEAQHYNTYTCKFTGCGKVYRSQGELEKHLDDHSTPPEKVLPPEAQLNSSGDSIQPSEVNQNTAENIEKERSMLPSENNIENSLLADRSDAWDKSKAESAVTKQDQISASELRQANGPLSNGLENPATTPLLQSSEVAVSIKVSLNQGIEDNFGKQENSTVEGSGEALVTDLHTPVEDTCNDLCHPGFQERKEQDCFNDAHVTQNSLVNSETLKIGDLTPQNLERQVNNLMTFSVQNQAAFQNNLPTSKFECGDNVKTSSNLYNLPLKTLESIAFVPPQSDLSNSLGTPSVPPKAPVQKFSCQVEGCTRTYNSSQSIGKHMKTAHPDQYAAFKMQRKSKKGQKANNLNTPNNGKFVYFLPSPVNSSNPFFTSQTKANGNPACSAQLQHVSPPIFPAHLASVSTPLLSSMESVINPNITSQDKNEQGGMLCSQMENLPSTALPAQMEDLTKTVLPLNIDSGSDPFLPLPAESSSMSLFPSPADSGTNSVFSQLENNTNHYSSQIEGNTNSSFLKGGNGENAVFPSQVNVANNFSSTNAQQSAPEKVKKDRGRGPNGKERKPKHNKRAKWPAIIRDGKFICSRCYRAFTNPRSLGGHLSKRSYCKPLDGAEIAQELLQSNGQPSLLASMILSTNAVNLQQPQQSTFNPEACFKDPSFLQLLAENRSPAFLPNTFPRSGVTNFNTSVSQEGSEIIKQALETAGIPSTFEGAEMLSHVSTGCVSDASQVNATVMPNPTVPPLLHTVCHPNTLLTNQNRTSNSKTSSIEECSSLPVFPTNDLLLKTVENGLCSSSFPNSGGPSQNFTSNSSRVSVISGPQNTRSSHLNKKGNSASKRRKKVAPPLIAPNASQNLVTSDLTTMGLIAKSVEIPTTNLHSNVIPTCEPQSLVENLTQKLNNVNNQLFMTDVKENFKTSLESHTVLAPLTLKTENGDSQMMALNSCTTSINSDLQISEDNVIQNFEKTLEIIKTAMNSQILEVKSGSQGAGETSQNAQINYNIQLPSVNTVQNNKLPDSSPFSSFISVMPTKSNIPQSEVSHKEDQIQEILEGLQKLKLENDLSTPASQCVLINTSVTLTPTPVKSTADITVIQPVSEMINIQFNDKVNKPFVCQNQGCNYSAMTKDALFKHYGKIHQYTPEMILEIKKNQLKFAPFKCVVPTCTKTFTRNSNLRAHCQLVHHFTTEEMVKLKIKRPYGRKSQSENVPASRSTQVKKQLAMTEENKKESQPALELRAETQNTHSNVAVIPEKQLVEKKSPDKTESSLQVITVTSEQCNTNALTNTQTKGRKIRRHKKEKEEKKRKKRKKPVSQSLPVSQSLEEFPTFPTRRYSPYYSPYRRPYPYRRC*VC*VHHQGC*FAAFTIQQNLILQGC*FAAFTIQQNLILH*H*YQAVYQAVH*KH*KSSDDLPAFSALPAFSAEEVVEEEEEESSEEAGKESEETETKQTLKEFRCQVSDCSRIFQAITGLIQHYMKLHEMTPEEIESMTASVDVGKFPCDQLECKSSFTTYLNYVVHLEADHGIGLRASKTEEDGVYKCDCEGCDRIYATRSNLLRHIFNKHNDKHKAHLIRPRRLTPGQENMSSKANQEKSKSKHRGTKHSRCGKEGIKMPKTKRKKKNNLENKNAKIVQIEENKPYSLKRGKHVYSIKARNDALSECTSRFVTQYPCMIKGCTSVVTSESNIIRHYKCHKLSKAFTSQHRNLLIVFKRCCNSQVKETSEQEGAKNDVKDSDTCVSESNDNSRTTATVSQKEVEKNEKDEMDELTELFITKLINEDSTSVETQANTSSNVSNDFQEDNLCQSERQKASNLKRVNKEKNVSQNKKRKVEKAEPASAAELSSVRKEEETAVAIQTIEEHPASFDWSSFKPMGFEVSFLKFLEESAVKQKKNTDKDHPNTGNKKGSHSNSRKNIDKTAVTSGNHVCPCKESETFVQFANPSQLQCSDNVKIVLDKNLKDCTELVLKQLQEMKPTVSLKKLEVHSNDPDMSVMKDISIGKATGRGQY

Zinc finger protein 292: 2723 aa, 305KD, pI 7.27, 67+

1

2

3

4

29

Region 1(1-680) 2(681-1361) 3(1362-2042) 4(2043-2733)

K/R/H +50/+28/+10 +50/+17/+21 +47/+19/+14 +83/+32/+23

K+R +78 +67 +66 +115

K+R+H +88 +88 +80 +138

K+R-D-E -29 -2 +10 +20

K+R+H-D-E -19 +19 +24 +43

-D-E -107 -69 -56 -95

-D/-E -34/-73 -27/-42 -16/-40 -30/-65

K+R+H+D+E 195 157 136 233

S*/T*Y*(total) 22/6/3(31) 27/6/2(35) 32/6/1(39) 31/13/2(46)

S/T/Y (total) 44/34/21(99) 74/38/14(126) 78/58/7(143) 67/47/14(128)

G/P/C/N/Q 26/28/28/26/40 35/46/19/62/43 24/45/16/60/44 23/21/23/42/35

pI 5.00 6.91 8.78 8.96

Aliphatic index 92.03 62.44 78.15 61.67

Hydropathicity -0.356 -0.692 -0.445 -0.491

Instability index 44.17 48.84 51.34 51.37