Oligosaccharyltransferase directly binds to ribosome at a location near the translocon-binding site

Oligosaccharyltransferase (OT) transfers high mannose-type glycans to the nascent polypeptides that are translated by the membrane-bound ribosome and translocated into the lumen of the endoplasmic reticulum through the Sec61 translocon complex. In this article, we show that purified ribosomes and OT...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2009-04, Vol.106 (17), p.6945-6949
Main Authors: Harada, Yoichiro, Li, Hua, Li, Huilin, Lennarz, William J
Format: Article
Language:English
Subjects:
60 APPLIED LIFE SCIENCES
Antibodies
BASIC BIOLOGICAL SCIENCES
Binding Sites
Biocatalysis
Biochemistry
Biological Sciences
Biological Transport
Carbohydrates
Cell membranes
Centrifugation
Data processing
Detergents
electron microscopy
ELECTRONS
ENDOPLASMIC RETICULUM
glycoprotein biosynthesis
Glycoproteins
Glycosylation
Hexosyltransferases - genetics
Hexosyltransferases - isolation & purification
Hexosyltransferases - metabolism
Hexosyltransferases - ultrastructure
IN VITRO
Membrane Proteins - genetics
Membrane Proteins - isolation & purification
Membrane Proteins - metabolism
Membrane Proteins - ultrastructure
Membrane Transport Proteins
Microscopy, Electron
multicomponent complexes
Phosphatidylcholines
POLYPEPTIDES
Protein Binding
Protein transport
Ribonucleic acid
RIBOSOMES
Ribosomes - metabolism
Ribosomes - ultrastructure
RNA
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - metabolism
Saccharomyces cerevisiae - ultrastructure
Saccharomyces cerevisiae Proteins - genetics
Saccharomyces cerevisiae Proteins - metabolism
SEC Translocation Channels
STOICHIOMETRY
TRANSLOCATION
Yeasts
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Summary:Oligosaccharyltransferase (OT) transfers high mannose-type glycans to the nascent polypeptides that are translated by the membrane-bound ribosome and translocated into the lumen of the endoplasmic reticulum through the Sec61 translocon complex. In this article, we show that purified ribosomes and OT can form a binary complex with a stoichiometry of [almost equal to]1 to 1 in the presence of detergent. We present evidence that OT may bind to the large ribosomal subunit near the site where nascent polypeptides exit. We further show that OT and the Sec61 complex can simultaneously bind to ribosomes in vitro. Based on existing data and our findings, we propose that cotranslational translocation and N-glycosylation of nascent polypeptides are mediated by a ternary supramolecular complex consisting of OT, the Sec61 complex, and ribosomes.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0812489106