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PMID: 3053734 Published · ppublish English Journal Article

Topogenic analysis of the human immunodeficiency virus type 1 envelope glycoprotein, gp160, in microsomal membranes.

The Journal of cell biology ·Vol. 107 ·No. 5 ·1988-11-00 ·Pages 1677-87

Haffar OK, Dowbenko DJ, Berman PW

Abstract

The orientation in cellular membranes of the 856 amino acid envelope glycoprotein precursor, gp160, of human immunodeficiency virus type 1 was investigated in vitro. Variants of the env gene were transcribed using the bacteriophage SP6 promoter, translated using a rabbit reticulocyte lysate, and translocated into canine pancreatic microsomal membranes. Immunoprecipitation studies of gp160 variants using antibodies specific for various gp160-derived polypeptides provided evidence that the external (cell surface) domain of gp160 begins at the mature amino terminus of the protein and continues through amino acid 665. A stop-transfer sequence (transmembrane domain) was identified in a hydrophobic region COOH-terminal to amino acid 665 and NH2-terminal to amino acid 732. Protease protection experiments demonstrated that gp160 possesses a single cytoplasmic domain COOH-terminal to residue 707. Membrane extraction studies using carbonate buffer provided evidence that the 29 amino acid hydrophobic domain (residues 512-541) of gp160 was unable to serve as a stop-transfer sequence. Finally, we propose that the cytoplasmic tail of gp160 forms a secondary association with the microsomal membranes.

MeSH Terms
Animals Antibody Specificity Biological Transport Cloning, Molecular Electrophoresis, Polyacrylamide Gel Endopeptidases/pharmacology HIV/genetics,physiology HIV Antibodies/immunology HIV Envelope Protein gp160 In Vitro Techniques Intracellular Membranes/analysis Microsomes/analysis Models, Biological Plasmids Precipitin Tests Protein Conformation Protein Precursors/analysis Recombinant Fusion Proteins/analysis,genetics Retroviridae Proteins/analysis,genetics Sulfur Radioisotopes Terminology as Topic Transcription, Genetic Viral Envelope Proteins/analysis,genetics
Chemicals
HIV Antibodies HIV Envelope Protein gp160 Protein Precursors Recombinant Fusion Proteins Retroviridae Proteins Sulfur Radioisotopes Viral Envelope Proteins Endopeptidases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Haffar O K
Department of Molecular Biology, Genentech, Inc., South San Francisco, California 94080.
Dowbenko D J
Berman P W
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.107.5.1677
Published
1988-11-00
Pages
1677-87
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2115321
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.107.5.1677 PMC Full Text View journal details
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