期刊
BIOCHEMISTRY
卷 52, 期 37, 页码 6487-6498出版社
AMER CHEMICAL SOC
DOI: 10.1021/bi400925g
关键词
-
资金
- LVMH-Recherche (Saint Jean de Braye, France)
- Institut National de la Sante et de la Recherche Medicale (INSERM)
- Association Nationale de la Recherche Technique (ANRT)
- Agence Nationale de la Recherche (WallArrayII Project) [ANR-10-GENM-0010]
- German Research Council [DFG SFB-TRR67 TPA7]
- Studienstiftung des deutschen Volkes
- Agence Nationale de la Recherche (ANR) [ANR-10-GENM-0010] Funding Source: Agence Nationale de la Recherche (ANR)
Human cysteine cathepsin S (catS) participates in distinct physiological and pathophysiological cellular processes and is considered as a valuable therapeutic target in autoimmune diseases, cancer, atherosclerosis, and asthma. We evaluated the capacity of negatively charged glycosaminoglycans (heparin, heparan sulfate, chondroitin 4/6-sulfates, dermatan sulfate, and hyaluronic acid) to modulate the activity of catS. Chondroitin 4-sulfate (C4-S) impaired the collagenolytic activity (type IV collagen) and inhibited the peptidase activity (Z-Phe-Arg-AMC) of catS at pH 5.5, obeying a mixed-type mechanism (estimated K-i = 16.5 +/- 6 mu M). Addition of NaCl restored catS activity, supporting the idea that electrostatic interactions are primarly involved. Furthermore, C4-S delayed in a dose-dependent manner the maturation of procatS at pH 4.0 by interfering with the intermolecular processing pathway. Binding of C4-S to catS was demonstrated by gel-filtration chromatography, and its affinity was measured by surface plasmon resonance (equilibrium dissociation constant K-d = 210 +/- 40 nM). Moreover, C4-S induced subtle conformational changes in mature catS as observed by intrinsic fluorescence spectroscopy analysis. Molecular docking predicted three specific binding sites on catS for C4-S that are different from those found in the crystal structure of the cathepsin K-C4-S complex. Overall, these results describe a novel glycosaminoglycan-mediated mechanism of catS inhibition and suggest that C4-S may modulate the collagenase activity of catS in vivo.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据