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.