Thermal stability of calf skin collagen type I in salt solutions

Komsa-Penkova, Regina; Koynova, Rumiana; Kostov, G.; Tenchov, Boris

doi:10.1016/s0167-4838(96)00092-1

Cited by 111 publications

(88 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…6A). This suggests that binding interactions between collagen and GAGs as described by Komsa-Penkova et al (36) do not significantly stabilize or destabilize the triple helical structure of the protein. However, the 1 ⁄4 and 3 ⁄4 polypeptide fragments of collagen generated by MMP-1 have significantly lower thermal stabilities than the parent protein.…”

Section: Degradation Of Type I and Ii Collagens By Lysosomal Cathepsimentioning

confidence: 69%

Regulation of Collagenase Activities of Human Cathepsins by Glycosaminoglycans

Yasuda

et al. 2004

Journal of Biological Chemistry

210

View full text Add to dashboard Cite

Cathepsin K, a lysosomal papain-like cysteine protease, forms collagenolytically highly active complexes with chondroitin sulfate and represents the most potent mammalian collagenase. Here we demonstrate that complex formation with glycosaminoglycans (GAGs) is unique for cathepsin K among human papain-like cysteine proteases and that different GAGs compete for the binding to cathepsin K. GAGs predominantly expressed in bone and cartilage, such as chondroitin and keratan sulfates, enhance the collagenolytic activity of cathepsin K, whereas dermatan, heparan sulfate, and heparin selectively inhibit this activity. Moreover, GAGs potently inhibit the collagenase activity of other cysteine proteases such as cathepsins L and S at 37°C. Along this line MMP1-generated collagen fragments in the presence of GAGs are stable against further degradation at 28°C by all cathepsins but cathepsin K, whereas thermal destabilization at 37°C renders the fragments accessible to all cathepsins. These results suggest a novel mechanism for the regulation of matrix protein degradation by GAGs. It further implies that cathepsin K represents the only lysosomal collagenolytic activity under physiologically relevant conditions.Controlled degradation of collagen is observed in bone remodeling, wound healing, angiogenesis, and during organ development (1-3). On the other hand excessive collagen degradation leads to pathological phenotypes such as osteoporosis (4), various forms of arthritis (5), or aneurysms of blood vessels (6), or it is characteristic for tumor invasion (7). However, triple helical collagens, in particular type I and II collagens, are highly resistant to general proteolysis and require specific proteases for their degradation. Known mammalian collagenolytic activities include members of the matrix metalloprotease family such as MMP-1, -2, -8, -13, and -14 (8), the serine protease, human neutrophil elastase (9), and thiol-dependent cathepsins (1). Collagenases of the MMP family cleave triple helical collagen at a specific single site and release 3 ⁄4 and 1 ⁄4 fragments. Similar to MMPs, human neutrophil elastase generates 3 ⁄4 fragments from type I collagen but is unable to degrade type II collagen (9). Lysosomal cysteine proteases such as cathepsins L and B have also been discussed as collagenolytic activities, but these data were mostly based on inhibitor experiments in cell extracts or on early preparations of cathepsins, which may not have excluded contaminating activities (10 -13). Thorough enzymatic studies suggested that cathepsins B and L primarily cleave in the non-helical telopeptide extensions of collagens (14, 15). A truly triple helical collagenase activity is found in cathepsin K, which is predominantly expressed in osteoclasts and to a lower degree in various other cell types including fibroblasts (16 -19). It was demonstrated that cathepsin K, similar to the bacterial Clostridium collagenase, cleaves at multiple sites within the triple helical region of types I and II collagens (20,21). The biological relevan...

show abstract

Section: Degradation Of Type I and Ii Collagens By Lysosomal Cathepsimentioning

confidence: 69%

Regulation of Collagenase Activities of Human Cathepsins by Glycosaminoglycans

Yasuda

et al. 2004

Journal of Biological Chemistry

210

View full text Add to dashboard Cite

show abstract

“…It should be noted that the inhibition of collagen degradation in the presence of 300 mM NaCl can not be attributed to a stabilization of the collagen structure by salt because, on the contrary, NaCl slightly destabilizes type I collagen (20). Similarly, the absence of chondroitin sulfate also prevents the collagenase activity of cathepsin K. This suggests that monomeric cathepsin K and cathepsin K in the absence of C4-S has no or only a minimal collagenase activity, which is highly reminiscent of the lack of collagenase activities in the catalytic domains of MMP collagenases (21,22).…”

Section: Discussionmentioning

confidence: 99%

“…The absorbance data were acquired at 280 nm as an average of 10 measurements at each radial position in 0.003-cm increments. A total of six data sets were collected corresponding to three different concentrations (10,20, and 50 M of cathepsin K and C4-S in 100 mM sodium acetate buffer, pH 5.5, containing 1 mM DTT, 1 mM EDTA, and 80 mM NaCl) and two speeds (6000 and 8000 rpm). Data editing and evaluation of the equilibrium were performed using the programs REEDIT and MATCH, respectively (provided by the National Analytical Ultracentrifugation facility of the University of Connecticut).…”

Section: Methodsmentioning

confidence: 99%

Collagenase Activity of Cathepsin K Depends on Complex Formation with Chondroitin Sulfate

Li¹,

Hou²,

Escalante-Torres³

et al. 2002

Journal of Biological Chemistry

160

141

View full text Add to dashboard Cite

Bone resorption in balance with bone formation is vital for the maintenance of the skeleton and is mediated by osteoclasts. Cathepsin K is the predominant protease in osteoclasts that degrades the bulk of the major bone forming organic component, type I collagen. Although the potent collagenase activity of cathepsin K is well known, its mechanism of action remains elusive. Here, we report a cathepsin K-specific complex with chondroitin sulfate, which is essential for the collagenolytic activity of the enzyme. The complex is an oligomer consisting of five cathepsin K and five chondroitin sulfate molecules. Only the complex exhibits potent triple helical collagen-degrading activity, whereas monomeric cathepsin K has no collagenase activity. The primary substrate specificity of cathepsin K is not altered by complex formation, suggesting that the protease-chondroitin sulfate complex primarily facilitates the destabilization and/or the specific binding of the triple helical collagen structure. Inhibition of complex formation leads to the loss of collagenolytic activity but does not impair the proteolytic activity of cathepsin K toward noncollagenous substrates. The physiological relevance of cathepsin K complexes is supported by the findings that (i) the content of chondroitin sulfate present in bone and accessible to cathepsin K activity is sufficient for complex formation and (ii) Y212C, a cathepsin K mutant that causes pycnodysostosis (a bone sclerosing disorder) and that has no collagenase activity but remains potent as a gelatinase, is unable to form complexes. These findings reveal a novel mechanism of bone collagen degradation and suggest that targeting cathepsin K complex formation would be an effective and specific treatment for diseases with excessive bone resorption such as osteoporosis.

show abstract

“…Nonetheless, it should be another important factor since the dielectric constant of aqueous medium is affected by the addition of salts. On this note, it was reported that many types of salts lowered the Td of CSC at a low concentration, while they raised the Td at higher concentration by salting out 12) . In the present study, neither buffers nor salts were used to adjust and stabilize the pH of each solution.…”

Section: Stability Index Of Collagenmentioning

confidence: 96%

New Index for the Stability of a Type I Collagen Affected by Hydrophobic Environment

et al. 2007

View full text Add to dashboard Cite

Effects of hydrophobic environment adjusted by various alcohols on the structural stability of calfskin collagen (CSC) were studied to elucidate the nature of collagen-monomer interaction in adhesion. The stability of CSC in aqueous alcohol solutions was represented by its denaturation temperature, Td, measured by DSC. The hydrophobicity of the alcohol solutions was quantified with their specific dielectric constants, εr, calculated from their concentrations. The effect of each alcohol to stabilize or destabilize CSC was evaluated by the initial slope of each Td vs. εr plot, denoted as -(dTd/dεr)ini and termed as stabilization power. Results showed that a hydrophobic environment with a smaller εr lowered the stabilization power. Stabilization power ranged from -3 (strong destabilization) for phenol (εr＝12) to ＋0.3 (weak stabilization) for glycerol (εr＝47). In view of the encouraging results obtained in this study, the new index was therefore helpful in predicting the effects of new dental materials of known εr values on the stability of dentinal collagen.

show abstract

Thermal stability of calf skin collagen type I in salt solutions

Cited by 111 publications

References 39 publications

Regulation of Collagenase Activities of Human Cathepsins by Glycosaminoglycans

Regulation of Collagenase Activities of Human Cathepsins by Glycosaminoglycans

Collagenase Activity of Cathepsin K Depends on Complex Formation with Chondroitin Sulfate

New Index for the Stability of a Type I Collagen Affected by Hydrophobic Environment

Contact Info

Product

Resources

About