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第六章 细胞内膜系统 与蛋白质分选. INTRACELLULAR COMPARTMENT AND PROTEINS SORTING. 本章内容提要. 第一节 蛋白质分选的基本原理 一、蛋白质分选信号 二、蛋白质分选运输的途径 第二节 膜泡运输 一、衣被类型 二、衣被的形成 三、膜泡运输的定向机制 四、细胞的内吞与外排. 第三节 内质网 一、形态与组成 二、 RER 的功能 三、 ER 的其它功能 第四节 高尔基体 一、形态与组成 二、功能区隔 三、主要功能 第五节 溶酶体与过氧化物酶体. - PowerPoint PPT Presentation
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INTRACELLULAR COMPARTMENT AND PROTEINS SORTING
RERER
,:? ? ?
signal sequencesorting receptor
signal sequence15-60signal peptidasesignal patch
signal sequence and signal patch
-Pro-Pro-Lys-Lys-Lys-Arg-Lys-Val-
-Leu-Ala-Leu-Lys-Leu-Ala-Gly-Leu-Asp-Ile-
+H3N-Met-Leu-Ser-Leu-Arg-Gln-Ser-Ile-Arg-Phe-Phe-Lys-Pro-Ala-Thr-Arg
-Thr-Leu-Cys-Ser-Ser-Arg-Tyr-Leu-Leu-
+H3N-Met-Val-Ala-Met-Ala-Met-Ala-Ser-Leu-Gln-Ser-Ser-Met-Ser-Ser-Leu-
Ser-Leu-Ser-Ser-Asn-Ser-Phe-Leu-Gly-Gln-Pro-Leu-Ser-Pro-Ile-Thr-Leu-
Ser-Pro-Phe-Leu-Gln-Gly-
-Ser-Lys-Leu-COO-
+H3N-Met-Met-Ser-Phe-Val-Ser-Leu-Leu-Leu-Val-Gly-Ile-Leu-Phe-Trp-Ala-
Thr-Glu-Ala-Glu-Gln-Leu-Thr-Lys-Cys-Glu-Val-Phe-Gln-
-Lys-Asp-Glu-Leu-COO-
1gated transport2transmembrane transporttranslocator3vesicular transport
coat
3dyneinkinesinmyosin
clathrinCOPICOPII
KDEL, KDELCOP,
(),(adapter),(dynamin)GTP
50100nm180kDa3540kDa, --(triskelion), (three-legged protein),, (a) ;(b);(c) P401
(adaptin):AP1AP2AP3AP1: AP1M6P,AP1AP2AP2: AP2()()AP3: , AP3AP3,
Clathrin coated vesicles
, (), , ,
, , , , , Hsp70, Ca2+ , Ca2+ , Ca2+ , , Ca2+ ,
dynamin1.5nmpinch off,900, GTPGTP,GTP,
COP I ( COP I ) COP,7:COP, COPCOPCOP escaped proteins
Cop I Vesicles
Cop I and II VesiclesCOP I
Lys-Asp-Glu-LeuKDELLys-Asp-Glu-LeuKDEL
SRPCLys-Lys-X-XCOP
COP GTP,ARF,GDP,GTP,ARF-GTP,GDPGTP ARFGTPaseARFGDP,GTPGTPARFARFGTP COPARF CoA(fatty-acyl CoA)COP,CoACOPCOP, , ARFGTP
COPSar1GTPSec23/Sec24Sec13/Sec31exit sitesCOP IICOP II[DE]X[DE] Asp-X-Glu COP
GTPcoat-recruitment GTPaseGTPmonomeric GTPaseGguanine-nucleotide exchange factor, GEFGDPGTP GTPGTPase activating protein, GAPGTP GTPARFSAR 1ARFCOP ISAR 1COP II
GTPGDPCOPIISAR1GDPGTPSAR 1SAR1Dphospholipase D
Coat assemblyCOPSar1GSar1GDPGTP, Sec23Sec24,Sec13Sec31,ER,Sec16Sec23/Sec24Sec13/Sec31,COP
, ???
: SNARE soluble NSF attachment protein receptorJames Rothman,N-(N-ethylmaleimide-sensitive fusion protein,NSF)NSF(soluble NSF attachment protein,SNAPs)NSFSNAPs --- SNAPsSNARE NSF/ SNAPs,Rothman:SNAP(SNAP receptors)SNAREsSNAPs
SNAREs20SNAREsv-SNAREst-SNAREs
v-t-SNAREsSNAREstrans-SNAREs complexes
SNAREs in vesicle transportRothmanSNAREV-SNARET-SNARE V-SNARE, T-SNARE,
NSF SNAREsATPadaptor proteinSNAREs
SNAREsSNAREsSNAREs
HIV fusion protein
Rabs RabGTP Rabstargeting GTPaseGTP30RabsRabsRabsGTPGDPGTPSNAREsRabseffector Rab
Rabs in docking
Rab
Bulk-phase endocytosis(Receptor mediated endocytosis, RME)caveolae
coated pitsmolecular filter2%4Tyr-X-X-XPheLeuMetdynamin
Clathrin coated piton the cytosolic face of a cell
(pit); ; , , ; ,
,(), : ,, , : ,(M6PLDL),, ,, :
LDL ParticleLDL
LDL22nm, 1500800500,LDL55 kDaB100(apolipoprotein B-100), (a)LDL, apo-B,LDL(b)
LDL endocytosis(low-density lipoprotein, LDL) LDLLDLLDL
Recycling of LDL receptors
(1) LDL;(2); (3);(4)();(5)pH,LDL(); (6) ,; (7),;(8)LDL, (10),,LDL
, , , ()(apotransferrin), Fe3+, (ferrotransferrin),pH, ,, , , , ,, , , ,
LDL Endocytosis
Transcytosis ,, , ,, , pH=6,()pH=7,FcpH=6IgGFc, pH=7Fc-IgG,IgG,
SNARERabLDL
constitutive exocytosis pathwayTGNdefault pathwayregulated exocytosis pathway
K. R. Porter1945endoplasmic reticulumER,
rough endoplasmic reticulumRERsmooth endoplasmic reticulumSERRER , SER,, RERSERER
,
ER
RER
SER
ERERER60%40%ER3030-6-ribophorinRERP450SER
(microsome) ,,
ER-----
, , ,
----------- George Palade,(membrane-bounded ribosome), :,,, Colvin Redman David SabatiniRER(),
RER,N-,;(b)RER,RER,
G. Blobel1975(Signal hypothesis)N1999signal peptideN16~306-15start transfer sequence,
AA Sequences of ER Signal Peptides : +H3N ; COO
1975, G.Blobel B.Dobberstein ,(signal hypothesis),(1);(2)N-,,N-;(3),;(4),,,,
signal recognition particleSRP67S RNARNP(ribonucleoprotein)[]SRPSPR receptorSRPSRP, stop transfer sequencePrtranslocatortranslocon3-4Sec61Sec613
Translocation of soluble proteins across ER
ERERmRNASRPSRP; ,SRPA,
SRP(), (protein-translocating channel) SRP SRP---mRNA,SRPGTPSRP,,,SRP,SRP,(),,,
,(signal peptidase),,
SRPSRPSRP co-translation
GTP SRPSRPG,, GTP-GDP, (translocon), GTP
(internal signal peptides),N-,,SRP,,,,, SRP,,,,(a) ,,(b)(a),,,
Insertion of a Multipass Transmembrane protein into the ER membrane
-----
42O-O-linked glycosylationSerThrHypOHN-N-N-linked glycosylationNH2N-
N-CMP-GDP-UDP-N-dolichol phosphateAsn-X-SerAsn-X-Thr
Protein glycosylation in RER
ERER,ERERER,ER,
COP IIexit siteshsp70ATP90%T
BipBip (heavy-chain binding protein) ,IgGBip,Hsp70,BipBipDNA,BipBipERBipERER
PTPP4506-
, -6-,-1-,-6-,,-6--6-,
,,,CDP-,
(1)P-450;(2)P-450NADPH; (3)P-450;(4),
18551889Golgi2050
cis facetrans face1um4~8()(Golgi stack)
The Golgi Apparatus
60%40%ERER-
cis Golgi networkCGNmedial Golgitrans Golgi networkTGN
cis(TPP)trans12(NADP)(CMP)transCMP
, , , 1RERERCOPIICGNmedial GdgiTGN
ER
:Lys-Asp-Glu-Leu-COO-KDELKDELERER, ER,, ER""ER
ER,: , ER ; ,,, ;,(4a)(4b);,(6a)(6b)
(constitutive secretory pathway), , (transport vesicles)(regulated secretory pathway), , ,, (secretory vesicles):;,200
2O-
, , 3 RER , 5, 2N-22,
34NCC56
B, N- , (preproinsulin), 12,000, , (proinsulin),9,000, 84, , 51C
KDEL,
N-O-
lysosomeC. de DuveB. Novikoff 1955
H+PH
0.2~0.5um60PH51primary lysosome
2Secondary lysosomeautophagolysosome(phagolysosome)
3residual body post-lysosome
10
, ,, ,
6-(mannose 6-phosphate, M6P)transN-cis1~2N-M6Ptransclathrin
(signal patch) , , , ,
N- N-, N- N-,N-UDP-GlcNAc
UDP N-(N-acetyglucosamineGlcNAc),
Transport of newly synthesized hydrolases to lysosomes(early endosome)(late endosome), pH, ATPase-H+ H+ pH
, ,M6P,, , pH, M6P, . , , , M6P, , ,
, , , ,
3-Tay-Sachs dieseaseAGM2 IIPompe-1,4-Gaucher- inclusion-cell diseaseN-M6P
-
1998
, 19143,.194319501980 19781980819922319801630 4CytomixisB 1998ProtoplasmaI19871985198621999
Rhodin 19540.5umflavin40H2O2RH2+O2R+H2O2
Peroxisome of hepatocyte
- H2O21/4-A
-Ser-Lys-Leu-COO-PTP
1 2 3 4 forming facecis face trans face 1 AM6P B C D2 A B C D
3Following organelles are all surrounded by double bio-membranes except AMitochondrion BChloroplast. CER. DNucleus. 4clathrin____ A. (endosome) B. C. D. E. 5. ____ A. B. C. D. E.
6. v-SNAREs____ A. B. C. D. 7COP A. B. C. D.