Action of rat liver cathepsin L on collagen and other substrates.

Kirschke, Heidrun; Kembhavi, A A; Bohley, Peter; Barrett, Alan J.

doi:10.1042/bj2010367

Cited by 218 publications

(136 citation statements)

References 22 publications

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“…The rabbit liver isoenzymes isolated in this study are chromatographically similar to rat liver cathepsin L which is also highly active against Z-PheArg-NMec [1,6]. The collagenolytic cathepsin isolated from bovine spleen, cathepsin N, has been shown to elute from CM-Sephadex at a much lower salt concentration, similar to that which elutes rabbit liver cathepsin B [2].…”

Section: Resultssupporting

confidence: 51%

“…We have shown that the collagenolytic activity of bovine spleen was caused by cathepsin B and cathepsin N [5]. In [6], cathepsin L isolated from rat liver was also reported to degrade collagen. We have now investigated the cysteine proteinases of rabbit liver, with particular emphasis on the major sources of collagenolytic activity in this tissue.…”

Section: Introductionmentioning

confidence: 99%

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Purification and characterisation of collagenolytic cathepsins from rabbit liver

1982

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Section: Resultssupporting

confidence: 51%

Section: Introductionmentioning

confidence: 99%

Purification and characterisation of collagenolytic cathepsins from rabbit liver

1982

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“…After 5, 10 and 20 min at room temperature the same volume of 2% azocasein in 6 M urea was added and the assay was incubated 30 min at 40°C to determine the residual activity of the enzyme those of cathepsin S. The data are only valid for S from beef spleen with regard to the substrate substrate concentrations of 10 mM for Bz-Argspecificity with low-M, substrates (table 2). On the NH2 (K,,, of 3 mM has been determined for rat other hand, the same split position between Met liver cathepsin L [5]) and 10pM for Z-Phe-Argand Arg in the hexapeptide Leu-Trp-Met-Arg-PheNMec (K,,, of 7 ,uM has been determined for Val has been detected for cathepsin L from rat cathepsin L from rat [9]). …”

Section: Resultsmentioning

confidence: 57%

“…Cathepsins B and L were purified from rat liver lysosomes as in [5,9] and stored in 0.1 M acetate buffer (pH 5.0) containing 0.5 mM HgClz and 1 mM NazEDTA. Cathepsins B and S from beef spleen were purified as in [lo].…”

Section: Enzyme Preparationmentioning

confidence: 99%

Species variations amongst lysosomal cysteine proteinases

1984

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Properties of cathepsin L from rat liver lysosomes were compared with those of a similar enzyme, cathepsin S from beef spleen. Major characteristics of cathepsin L are the high activity against Z-Phe-Arg-methylcoumarylamide and sensitivity to the fast reacting irreversible inhibitor Z-Phe-Phe-diazomethane.In contrast, cathepsin S hydrolyzes Z-Phe-Arg-methylcoumarylamide only slowly and Z-Phe-Phe-diazomethane cannot be regarded as a potent inhibitor of this enzyme. The differences in the substrate specificity of cathepsin L from rat liver and cathepsin S from beef spleen are discussed in comparison with the substrate specificity of cathepsin B from rat and human liver and beef spleen.

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“…MMPs selectively cleave collagens through a single scission across all three chains and generate about three-fourth and one-fourth length collagen fragments (5) whereas cathepsin K, similar to bacterial collagenases, cleaves type I and II collagens at multiple sites within their triple helical domains (6,7). In contrast, other cysteine proteases such as cathepsins L and B cleave collagens only in their nonhelical telopeptide regions (8).…”

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confidence: 99%

Selective Inhibition of the Collagenolytic Activity of Human Cathepsin K by Altering Its S2 Subsite Specificity

et al. 2002

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The primary specificity of papain-like cysteine proteases (family C1, clan CA) is determined by S2-P2 interactions. Despite the high amino acid sequence identities and structural similarities between cathepsins K and L, only cathepsin K is capable of cleaving interstitial collagens in their triple helical domains. To investigate this specificity, we have engineered the S2 pocket of human cathepsin K into a cathepsin L-like subsite. Using combinatorial fluorogenic substrate libraries, the P1-P4 substrate specificity of the cathepsin K variant, Tyr67Leu/Leu205Ala, was determined and compared with those of cathepsins K and L. The introduction of the double mutation into the S2 subsite of cathepsin K rendered the unique S2 binding preference of the protease for proline and leucine residues into a cathepsin L-like preference for bulky aromatic residues. Homology modeling and docking calculations supported the experimental findings. The cathepsin L-like S2 specificity of the mutant protein and the integrity of its catalytic site were confirmed by kinetic analysis of synthetic di-and tripeptide substrates as well as pH stability and pH activity profile studies. The loss of the ability to accept proline in the S2 binding pocket by the mutant protease completely abolished the collagenolytic activity of cathepsin K whereas its overall gelatinolytic activity remained unaffected. These results indicate that Tyr67 and Leu205 play a key role in the binding of proline residues in the S2 pocket of cathepsin K and are required for its unique collagenase activity.Type I collagen is the major component of the organic bone matrix which is continually degraded and resynthesized during the bone remodeling process (1). It consists of covalently cross-linked triple helices containing two R1(I) and one R2(I) chains. Large regions of the collagen chains are composed of repeating Gly-Pro-Xaa sequences, where Xaa is mainly proline and/or 4-trans-L-hydroxyproline (2). Degradation of the triple helical type I collagen is achieved by several members of the matrix metalloprotease family (MMPs) such as MMPs 1, 2, 8, and 13 (3) and the papainlike cysteine protease, cathepsin K (4). MMPs selectively cleave collagens through a single scission across all three chains and generate about three-fourth and one-fourth length collagen fragments (5) whereas cathepsin K, similar to bacterial collagenases, cleaves type I and II collagens at multiple sites within their triple helical domains (6, 7). In contrast, other cysteine proteases such as cathepsins L and B cleave collagens only in their nonhelical telopeptide regions (8).Cathepsin K is predominantly expressed in osteoclasts and has been implicated in bone resorption (9-11). The specific role of cathepsin K in bone resorption was demonstrated by the discovery that deficiency in cathepsin K activity causes an inherited autosomal recessive bone dysplasia, pycnodysostosis (12). Several cleavage sites of cathepsin K in the triple helical regions of type I and II collagens have been identified (6,...

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Action of rat liver cathepsin L on collagen and other substrates.

Cited by 218 publications

References 22 publications

Purification and characterisation of collagenolytic cathepsins from rabbit liver

Purification and characterisation of collagenolytic cathepsins from rabbit liver

Species variations amongst lysosomal cysteine proteinases

Selective Inhibition of the Collagenolytic Activity of Human Cathepsin K by Altering Its S2 Subsite Specificity

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