Expression of human cathepsin K in Pichia pastoris and preliminary crystallographic studies of an inhibitor complex

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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 Å X-ray data set has been collected on crystals belonging to space group P212121, 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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