Open AccessPurification, crystallization and preliminary X‐ray diffraction studies of the soluble domain of the oligosaccharyltransferase STT3 subunit from the thermophilic archaeon Pyrococcus furiosus
Author(s) -
Igura Mayumi,
Maita Nobuo,
Obita Takayuki,
Kamishikiryo Jun,
Maenaka Katsumi,
Kohda Daisuke
Publication year - 2007
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309107040134
Subject(s) - pyrococcus furiosus , thermophile , protein subunit , protein crystallization , crystallization , crystallography , chemistry , biochemistry , escherichia coli , archaea , enzyme , organic chemistry , gene
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 Å. 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.