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...

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

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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

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ME A3IAKYDFKATADDELSFKRGDILKVLNEECDQNWYKAELNGKDGFIPKNYIE54 MKP57H P59WFFGKIPRAKAEEMLSKQRHDGAFLIRESESAPGDFSLSVKFGNDVQHFKVLRDGAGKYFLWVVKFNSLNELVDYHRSTSVSRNQQIFLR149 DIEQVP155QQP158T Y160VQALFDFDPQEDGELGFRRGDFIHVMDNSDPNWWKGACHGQTGMFPRNYVTP212 VNRNV

15

MKPHPWFFGKIPRAKAEEMLSKQRHDGAFLIRESESAPGDFSLSVKFGNDVQHFKVLRDGAGKYFLWVVKFNSLNELVDYHRSTSVSRNQQIFLRDIEQVP

Tertiary structure of SH2

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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

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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

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MKPHPWFFGKIPR67AKAEEMLSKQRHDGAFLIR86ESESAPGDFSLSVKFGNDVQH107F108K109VLRDGAGKYFLWVVKFNSLNELVDYH135RSTSVSRNQQIFLRDIEQVP

R67

R86

H107 K109

pTyr binding pocket

F108

H135

unique binding pocket

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Grb2 binds glycerol

glycerol

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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

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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

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Sos-A: SPLLPK6LPPK10TYKRE

SH3:GSRRASVGSMEAIAKYDFKATAD23D24E25*LSFKRGDILK

VLNEECD42QNWYKAELNGKDFIPKNYIEMKPHPEFIVTD

--- D23D24E25*

+ K6

+ K10

- D42

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Sos-A: SP2L3L4P5KL7P8P9KTYKRE

SH3:GSRRASVGSMEAIAKY16*D17F18*K19A20TADDE*LS

FKRGDILKVLNEECDQNW*YKAELNGKDF55I56P57*KNY*IEMKPHPEFIVTD

P2L3L4P5

L7P8P9

Y16D17F18K19A20

F55I56P57

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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

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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%

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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

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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