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PMID: 17768359 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Purification, crystallization and preliminary X-ray diffraction studies of the soluble domain of the oligosaccharyltransferase STT3 subunit from the thermophilic archaeon Pyrococcus furiosus.

Acta crystallographica. Section F, Structural biology and crystallization communications ·Vol. 63 ·No. Pt 9 ·2007-09-01 ·Pages 798-801

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

Abstract

Oligosaccharyltransferase catalyzes the transfer of preassembled oligosaccharides onto asparagine residues in nascent polypeptide chains. The STT3 subunit is thought to bear the catalytic site. The C-terminal domain of the STT3 protein of Pyrococcus furiosus was expressed in Escherichia coli cells. STT3 protein prepared from two different sources, the soluble fraction and the inclusion bodies, produced crystals that diffracted to 2.7 A. During crystallization screening, cocrystals of P. furiosus STT3 with an E. coli 50S ribosomal protein, L7/L12, were accidentally obtained. This cross-species interaction is not biologically relevant, but may be used to design a built-in polypeptide substrate for the STT3 crystals.

MeSH Terms
Archaeal Proteins/chemistry,isolation & purification Bacterial Proteins/chemistry,isolation & purification,metabolism Campylobacter/enzymology Glycosylation Hexosyltransferases/chemistry,genetics,isolation & purification,metabolism Hot Temperature Membrane Proteins/chemistry,genetics,isolation & purification,metabolism Protein Folding Protein Subunits/chemistry,isolation & purification Pyrococcus furiosus/enzymology Recombinant Proteins/chemistry,isolation & purification,metabolism
Chemicals
Archaeal Proteins Bacterial Proteins Membrane Proteins Protein Subunits Recombinant Proteins Hexosyltransferases dolichyl-diphosphooligosaccharide - protein glycotransferase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Igura Mayumi
Division of Structural Biology, Medical Institute of Bioregulation, Kyushu University, Maidashi 3-1-1, Higashi-ku, Fukuoka 812-8582, Japan.
Maita Nobuo
Obita Takayuki
Kamishikiryo Jun
Maenaka Katsumi
Kohda Daisuke
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Article Info
Journal
Acta crystallographica. Section F, Structural biology and crystallization communications
Abbr.
Acta Crystallogr Sect F Struct Biol Cryst Commun
ISSN
1744-3091
DOI
10.1107/S1744309107040134
Published
2007-09-01
Epub
2007-00-31
Pages
798-801
Language
English
Region
England
NLM ID
101226117
PMCID
PMC2376324
Subset
IM
External Links
PubMed source DOI: 10.1107/S1744309107040134 PMC Full Text
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