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...
TRANSCRIPT
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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
14
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
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