Open AccessUse of novel selenomethionine‐resistant yeast to produce selenomethionyl protein suitable for structural analysis
Author(s) -
Kitajima Toshihiko,
Yagi Emi,
Kubota Tomomi,
Chiba Yasunori,
Nishikawa Satoshi,
Jigami Yoshifumi
Publication year - 2009
Publication title -
fems yeast research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 92
eISSN - 1567-1364
pISSN - 1567-1356
DOI - 10.1111/j.1567-1364.2009.00484.x
Subject(s) - biology , mutant , yeast , selenium , glycosylation , biochemistry , lysozyme , pichia pastoris , saccharomyces cerevisiae , protein biosynthesis , gene , chemistry , recombinant dna , organic chemistry
Yeast is widely used to determine the tertiary structure of eukaryotic proteins, because of its ability to undergo post‐translational modifications such as glycosylation. A mutant lacking S ‐adenosylmethionine synthesis has been reported as a suitable host for producing selenomethionine derivatives, which can help solve phase problems in protein crystallography. However, the mutant required external addition of S ‐adenosylmethionine for cell proliferation. Here, a selenomethionine‐resistant Pichia pastoris mutant that showed S ‐adenosylmethionine autotrophy was isolated. Human lysozyme expressed by the mutant under the control of constitutive promoter contained selenomethionine at 65% occupancy, sufficient for use as a selenomethionine derivative for single‐wavelength anomalous dispersion phasing.