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Different Cysteine Proteinases Involved in Bone Resorption and Osteoclast Formation

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Abstract

Cysteine proteinases, especially cathepsin K, play an important role in osteoclastic degradation of bone matrix proteins and the process can, consequently, be significantly inhibited by cysteine proteinase inhibitors. We have recently reported that cystatin C and other cysteine proteinase inhibitors also reduce osteoclast formation. However, it is not known which cysteine proteinase(s) are involved in osteoclast differentiation. In the present study, we compared the relative potencies of cystatins C and D as inhibitors of bone resorption in cultured mouse calvariae, osteoclastogenesis in mouse bone marrow cultures, and cathepsin K activity. Inhibition of cathepsin K activity was assessed by determining equilibrium constants for inhibitor complexes in fluorogenic substrate assays. The data demonstrate that whereas human cystatins C and D are equipotent as inhibitors of bone resorption, cystatin D is 10-fold less potent as an inhibitor of osteoclastogenesis and 200-fold less potent as an inhibitor of cathepsin K activity. A recombinant human cystatin C variant with Gly substitutions for residues Arg8, Leu9, Val10, and Trp106 did not inhibit bone resorption, had 1,000-fold decreased inhibitory effect on cathepsin K activity compared to wildtype cystatin C, but was equipotent with wildtype cystatin C as an inhibitor of osteoclastogenesis. It is concluded that (i) different cysteine proteinases are likely to be involved in bone resorption and osteoclast formation, (ii) cathepsin K may not be an exclusive target enzyme in any of the two systems, and (iii) the enzyme(s) involved in osteoclastogenesis might not be a typical papain-like cysteine proteinase.

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Acknowledgment

This project was supported by a grant from the “Network for Inflammation Research” funded by the Swedish Foundation for Strategic Research and by grants from the Swedish Science Council (project no.07525, 05196, 09915), the Swedish Rheumatism Association, the Royal 80 Year Found of King Gustav V, A. Påhlsson’s, A. Österlund’s and G. & J. Kock’s Foundations, well as by the County Council of Västerbotten. The kind gift of cathepsin K from GlaxoSmithKline through the courtesy of Dr. Maxine Gowen is highly appreciated.

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Authors and Affiliations

  1. Department of Oral Cell Biology, Umeå university, s-90187 Umeå, Sweden

    M. Brage & U. H. Lerner

  2. Department of Clinical Chemistry, Institute of Laboratory Medicine, Lund university, S-22185 Lund, Sweden

    M. Abrahamson, V. Lindström & A. Grubb

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  1. M. Brage
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  2. M. Abrahamson
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Brage, M., Abrahamson, M., Lindström, V. et al. Different Cysteine Proteinases Involved in Bone Resorption and Osteoclast Formation. Calcif Tissue Int 76, 439–447 (2005). https://doi.org/10.1007/s00223-004-0043-y

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