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Expression of human cathepsin K in Pichia pastoris and preliminary crystallographic studies of an inhibitor complex
Author(s) -
Linnevers Christopher J.,
Mcgrath Mary E.,
Armstrong Andy,
Mistry Firoz R.,
Barnes Michael G.,
Klaus Jeffrey L.,
Palmer James T.,
Katz Bradley A.,
Brömme Dieter
Publication year - 1997
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.5560060421
Subject(s) - pichia pastoris , cathepsin e , cysteine protease , chemistry , biochemistry , cathepsin a , cathepsin h , cathepsin b , protease , glycosylation , cathepsin , mutant , cathepsin o , microbiology and biotechnology , biology , enzyme , recombinant dna , gene
Cathepsin K is a cysteine protease of the papain family, which is predominantly expressed in osteoclasts, and is regarded as a key protease in bone remodeling. To facilitate structural studies of the protein, the wild‐type sequence of the protease has been mutated so as to replace a potential N‐glycosylation site. We have expressed the mutant human cathepsin K to 190 mg/5 L using the Pichia pastoris expression system. Cathepsin K was inactivated with the mechanism‐based inhibitor, APC3328, and crystallized from magnesium formate. A 2.2 Å X‐ray data set has been collected on crystals belonging to space group P2 1 2 1 2 1 , with a = 41.66 Å, b = 51.41 Å, and c = 107.72 Å. There is most likely one molecule per asymmetric unit.
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