title: Expression of human cathepsin K in Pichia pastoris and preliminary crystallographic studies of an inhibitor complex.
abstract: 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 A X-ray data set has been collected on crystals belonging to space group P2(1)2(1)2(1), with a = 41.66 A, b = 51.41 A, and c = 107.72 A. There is most likely one molecule per asymmetric unit.
authors.affiliation: Arris Pharmaceutical, South San Francisco, California 94080, USA.
authors.forename: C J
authors.initials: CJ
authors.lastname: Linnevers
authors.forename: M E
authors.initials: ME
authors.lastname: McGrath
authors.forename: R
authors.initials: R
authors.lastname: Armstrong
authors.forename: F R
authors.initials: FR
authors.lastname: Mistry
authors.forename: M G
authors.initials: MG
authors.lastname: Barnes
authors.forename: J L
authors.initials: JL
authors.lastname: Klaus
authors.forename: J T
authors.initials: JT
authors.lastname: Palmer
authors.forename: B A
authors.initials: BA
authors.lastname: Katz
authors.forename: D
authors.initials: D
authors.lastname: Brömme
mesh_terms: D056657:Cathepsin K; D002403:Cathepsins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D004789:Enzyme Activation; D006801:Humans; D010843:Pichia / Q000201:enzymology*
chemical_list: D002403:Cathepsins; C534130:CTSK protein, human; D056657:Cathepsin K