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PMID: 3025652 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Determination of the orientation of an integral membrane protein and sites of glycosylation by oligonucleotide-directed mutagenesis: influenza B virus NB glycoprotein lacks a cleavable signal sequence and has an extracellular NH2-terminal region.

Molecular and cellular biology ·Vol. 6 ·No. 12 ·1986-12-00 ·Pages 4317-28

Williams MA, Lamb RA

Abstract

The membrane orientation of the NB protein of influenza B virus, a small (Mr, approximately 18,000) glycoprotein with a single internal hydrophobic domain, was investigated by biochemical and genetic means. Cell fractionation and protein solubility studies indicate NB is an integral membrane protein, and NB has been shown to be a dimer under nonreducing conditions. Treatment of infected-cell surfaces with proteinase K and endoglycosidase F and immunoprecipitation with a site-specific antibody suggests that the 18-amino-acid NH2-terminal region of NB is exposed at the cell surface. Oligonucleotide-directed mutagenesis to eliminate each of the four potential sites of N-linked glycosylation and expression of the mutant NB proteins in eucaryotic cells suggest that the two sites adjacent to the NH2 terminus are glycosylated. This provides further evidence that NB, which lacks a cleavable NH2-terminal signal sequence, has an exposed NH2 terminus at the cell surface.

MeSH Terms
Amino Acid Sequence Animals Cell Line Chick Embryo Clone Cells Dogs Influenza B virus/genetics Microsomes/metabolism Mutation Oligodeoxyribonucleotides/pharmacology Plasmids Simian virus 40/genetics Viral Proteins/genetics
Chemicals
NB glycoprotein, Influenza B virus Oligodeoxyribonucleotides Viral Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Williams M A
Lamb R A
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52 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.6.12.4317-4328.1986
Published
1986-12-00
Pages
4317-28
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC367213
Subset
IM
Grants
NIAID NIH HHS · AI-20201 · United States
NIAID NIH HHS · AI-23173 · United States
External Links
PubMed source DOI: 10.1128/mcb.6.12.4317-4328.1986 PMC Full Text View journal details
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