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PMID: 18046457 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Structure-guided identification of a new catalytic motif of oligosaccharyltransferase.

The EMBO journal ·Vol. 27 ·No. 1 ·2008-01-09 ·Pages 234-43

Igura M, Maita N, Kamishikiryo J, Yamada M, Obita T, Maenaka K, Kohda D

Abstract

Asn-glycosylation is widespread not only in eukaryotes but also in archaea and some eubacteria. Oligosaccharyltransferase (OST) catalyzes the co-translational transfer of an oligosaccharide from a lipid donor to an asparagine residue in nascent polypeptide chains. Here, we report that a thermophilic archaeon, Pyrococcus furiosus OST is composed of the STT3 protein alone, and catalyzes the transfer of a heptasaccharide, containing one hexouronate and two pentose residues, onto peptides in an Asn-X-Thr/Ser-motif-dependent manner. We also determined the 2.7-A resolution crystal structure of the C-terminal soluble domain of Pyrococcus STT3. The structure-based multiple sequence alignment revealed a new motif, DxxK, which is adjacent to the well-conserved WWDYG motif in the tertiary structure. The mutagenesis of the DK motif residues in yeast STT3 revealed the essential role of the motif in the catalytic activity. The function of this motif may be related to the binding of the pyrophosphate group of lipid-linked oligosaccharide donors through a transiently bound cation. Our structure provides the first structural insights into the formation of the oligosaccharide-asparagine bond.

MeSH Terms
Amino Acid Motifs Amino Acid Sequence Carbohydrate Sequence Catalytic Domain/genetics,physiology Crystallography, X-Ray Hexosyltransferases/chemistry,genetics,isolation & purification Membrane Proteins/chemistry,genetics,isolation & purification Molecular Sequence Data Oligosaccharides/chemistry,genetics Pyrococcus furiosus/enzymology,genetics Saccharomyces cerevisiae/enzymology,genetics
Chemicals
Membrane Proteins Oligosaccharides Hexosyltransferases dolichyl-diphosphooligosaccharide - protein glycotransferase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Igura Mayumi
Division of Structural Biology, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.
Maita Nobuo
Kamishikiryo Jun
Yamada Masaki
Obita Takayuki
Maenaka Katsumi
Kohda Daisuke
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
1460-2075
DOI
10.1038/sj.emboj.7601940
Published
2008-01-09
Epub
2007-00-29
Pages
234-43
Language
English
Region
England
NLM ID
8208664
PMCID
PMC2206122
Subset
IM
Databases
PDB
2ZAG, 2ZAI
External Links
PubMed source DOI: 10.1038/sj.emboj.7601940 PMC Full Text View journal details
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