Expression of human cathepsin K in Pichia pastoris and preliminary crystallographic studies of an inhibitor complex. |
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Authors: | C. J. Linnevers, M. E. McGrath, R. Armstrong, F. R. Mistry, M. G. Barnes, J. L. Klaus, J. T. Palmer, B. A. Katz, D. Br mme |
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Affiliation: | C. J. Linnevers, M. E. McGrath, R. Armstrong, F. R. Mistry, M. G. Barnes, J. L. Klaus, J. T. Palmer, B. A. Katz, and D. Brömme |
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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 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. |
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Keywords: | cysteine protease osteoclast osteopetrosis structure-based drug design vinyl sulfone |
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