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PMID:15308636

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Citation

Breuza, L, Halbeisen, R, Jenö, P, Otte, S, Barlowe, C, Hong, W and Hauri, HP (2004) Proteomics of endoplasmic reticulum-Golgi intermediate compartment (ERGIC) membranes from brefeldin A-treated HepG2 cells identifies ERGIC-32, a new cycling protein that interacts with human Erv46. J. Biol. Chem. 279:47242-53

Abstract

Cycling proteins play important roles in the organization and function of the early secretory pathway by participating in membrane traffic and selective transport of cargo between the endoplasmic reticulum (ER), the intermediate compartment (ERGIC), and the Golgi. To identify new cycling proteins, we have developed a novel procedure for the purification of ERGIC membranes from HepG2 cells treated with brefeldin A, a drug known to accumulate cycling proteins in the ERGIC. Membranes enriched 110-fold over the homogenate for ERGIC-53 were obtained and analyzed by mass spectrometry. Major proteins corresponded to established and putative cargo receptors and components mediating protein maturation and membrane traffic. Among the uncharacterized proteins, a 32-kDa protein termed ERGIC-32 is a novel cycling membrane protein with sequence homology to Erv41p and Erv46p, two proteins enriched in COPII vesicles of yeast. ERGIC-32 localizes to the ERGIC and partially colocalizes with the human homologs of Erv41p and Erv46p, which mainly localize to the cis-Golgi. ERGIC-32 interacts with human Erv46 (hErv46) as revealed by covalent cross-linking and mistargeting experiments, and silencing of ERGIC-32 by small interfering RNAs increases the turnover of hErv46. We propose that ERGIC-32 functions as a modulator of the hErv41-hErv46 complex by stabilizing hErv46. Our novel approach for the isolation of the ERGIC from BFA-treated cells may ultimately lead to the identification of all proteins rapidly cycling early in the secretory pathway.

Links

PubMed Online version:10.1074/jbc.M406644200

Keywords

Amino Acid Sequence; Animals; Brefeldin A/pharmacology; Carrier Proteins/biosynthesis; Carrier Proteins/chemistry; Cell Line; Cell Membrane/metabolism; Cross-Linking Reagents/pharmacology; Cytoplasm/metabolism; Electrophoresis, Polyacrylamide Gel; Endoplasmic Reticulum/metabolism; Golgi Apparatus/metabolism; HeLa Cells; Humans; Immunoblotting; Immunoprecipitation; Mass Spectrometry; Membrane Proteins/biosynthesis; Membrane Proteins/chemistry; Membrane Proteins/metabolism; Mice; Microscopy, Fluorescence; Models, Biological; Molecular Sequence Data; Phylogeny; Protein Binding; Protein Structure, Tertiary; Proteome/chemistry; RNA Interference; Saccharomyces cerevisiae Proteins/chemistry; Saccharomyces cerevisiae Proteins/metabolism; Sequence Homology, Amino Acid; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Subcellular Fractions/metabolism; Vesicular Transport Proteins/chemistry

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:SURF4

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:SC22B

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:YIF1A

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:O95330

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q969X5

F

Seeded From UniProt

complete

HUMAN:O95330

involved_in

GO:0006888: endoplasmic reticulum to Golgi vesicle-mediated transport

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:O95330

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:FINC

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:PDIA1

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:BIP

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:AMPN

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:ERD21

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MA1A1

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:LMAN1

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:SERPH

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:NUCB2

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:NUCB1

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:LMAN2

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:GOGB1

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:PDIA6

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:ERGI1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O95330

F

Seeded From UniProt

complete

HUMAN:ERGI1

involved_in

GO:0006888: endoplasmic reticulum to Golgi vesicle-mediated transport

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:ERGI1

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:RAB37

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:ERP44

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:Q9HC06

involved_in

GO:0006888: endoplasmic reticulum to Golgi vesicle-mediated transport

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:Q9HC06

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:TMED5

located_in

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MA1A1

part_of

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:PDIA6

part_of

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:SERPH

part_of

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:PDIA1

part_of

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:FINC

part_of

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:ERP44

part_of

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:BIP

part_of

GO:0005793: endoplasmic reticulum-Golgi intermediate compartment

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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