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PMID: 7729412 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Folding and oligomerization of influenza hemagglutinin in the ER and the intermediate compartment.

The EMBO journal ·Vol. 14 ·No. 7 ·1995-04-03 ·Pages 1340-8

Tatu U, Hammond C, Helenius A

Abstract

Influenza hemagglutinin (HA) was used to analyze the stepwise folding and oligomeric assembly of glycoproteins in the early secretory pathway of living cells. In addition to mature trimers, six distinct maturation intermediates were identified. Of these, all the incompletely oxidized forms were located in the endoplasmic reticulum (ER) and associated with calnexin, a membrane-bound, lectin-like ER chaperone. Once fully oxidized, the HA dissociated from calnexin as a monomer, which rapidly became resistant to dithiothreitol (DTT) reduction. Part of these extensively folded molecules moved as monomers into the intermediate compartment between the ER and the Golgi complex. Assembly of homotrimers occurred without calnexin-involvement within the ER and in the intermediate compartment. When anchor-free HA molecules were analyzed, it was found that they reach the DTT-resistant monomeric conformation but fail to trimerize. Taken together, the results provide a definition and intracellular localization of several intermediates in the conformational maturation of HA, including the immediate precursor for trimer assembly.

MeSH Terms
Animals CHO Cells Cell Fractionation Centrifugation, Density Gradient Clone Cells Cricetinae Dithiothreitol/pharmacology Endoplasmic Reticulum/metabolism Golgi Apparatus/metabolism Hemagglutinin Glycoproteins, Influenza Virus Hemagglutinins, Viral/chemistry,isolation & purification,metabolism Kinetics Macromolecular Substances Models, Structural Protein Conformation/drug effects Protein Folding Recombinant Proteins/chemistry,isolation & purification,metabolism Subcellular Fractions/metabolism Time Factors Viral Envelope Proteins/metabolism
Chemicals
Hemagglutinin Glycoproteins, Influenza Virus Hemagglutinins, Viral Macromolecular Substances Recombinant Proteins Viral Envelope Proteins Dithiothreitol
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Tatu U
Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA.
Hammond C
Helenius A
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1995-04-03
Pages
1340-8
Language
English
Region
England
NLM ID
8208664
PMCID
PMC398219
Subset
IM
Grants
NCI NIH HHS · 5PO1 CA 46182 · United States
NIGMS NIH HHS · 5RO1 GM 38346 · United States
External Links
PubMed source PMC Full Text View journal details
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