Cathepsin D: Cleavage of soluble collagen and crosslinked peptides

Scott, Paul; Pearson, C. H.

doi:10.1016/0014-5793(78)80602-4

Cited by 25 publications

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References 17 publications

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“…3 products when they assayed at pH 4.2 (30sC). Scott and Pearson (22,23) found that the cleavage of crosslinked bovine collagen by cathepsin D at pH 4.0 and 45cC occurred at position 6h-7e (Leu-Ser in calf) in the COOH telopeptide. * Consistent with this, we observed that cleavage of native procollagen in the pH range 2.5-5.0 at 37tC resulted in a cleavage product larger than the authentic C-i subunit by at least 1.2 kDa.…”

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Cathepsin D-mediated processing of procollagen: lysosomal enzyme involvement in secretory processing of procollagen.

Helseth¹,

Veis²

1984

Proc. Natl. Acad. Sci. U.S.A.

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The proteolytic removal of the extension COOH-terminal propeptide from procollagen has been examined in vitro. A crude enzyme activity was identified in a whole-chicken-embryo extract that acted at acid pH and appeared to be similar to one identified previously [Davidson, J. M., McEneany, L. S. G. & Bornstein, P. (1979) Eur. J. Biochem. 100,[551][552][553][554][555][556][557][558]. This activity was inhibitable by pepstatin but not by leupeptin, suggesting that it might be cathepsin D. Cathepsin D was purified 907-fold from chicken livers by affinity chromatography on pepstatin-aminohexyl-Sepharose 4B and was found to remove the COOH propeptides from procollagen. At pH 6.0, the site of cleavage appeared to shift from the COOH telopeptide to the COOH telopeptide/propeptide junction, based upon the difference in electrophoretic migration of the cleavage products, although determining the actual cleavage site will require end-group analysis. A model for the involvement of cathepsin D in the in vivo processing of procollagen is presented.The enzymes responsible for the sequential removal of the extension propeptides from procollagen, procollagen NH2-terminal proteinase and procollagen COOH-terminal proteinase, have been actively studied, but only the former has been well characterized (1). In vivo it appears that the COOH-terminal propeptide (COOH propeptide) is removed first, with the NH2-terminal propeptide (NH2 propeptide) removed later, possibly after the intermediate has assembled into extracellular fibrils (2-4). The site of COOH propeptide removal has not been well defined. In cell culture experiments, the apparent in vivo pathway is reversed. The COOH propeptide is not cleaved from the product that accumulates in cell culture media, yet conversion is complete in organ culture (5), suggesting that an intact extracellular matrix is required for conversion.In searching for a neutral metalloproteinase that is a COOH-terminal proteinase (6), we observed a cathepsin Dlike activity, which removed the COOH propeptide from procollagen at acid pH. We report here that the cleavage of procollagen with purified cathepsin D is specific for the COOH propeptide and that cleavage occurs at or near the authentic cleavage site when digestion is carried out near pH 6.0. We postulate a role for cathepsin D in the in vivo conversion of procollagen to collagen. MATERIALS AND METHODSProcollagen was isolated from 17-day chicken-embryo tendon fibroblasts (7,8). After tendon dissection and dissociation with collagenase and trypsin, fibroblasts were incubated (2 x 107 cells per ml) for 6 hr in modified Krebs medium II in the presence of 1 ,uCi (1 Ci = 37 GBq) of a mixture of 15 14C-labeled amino acids (New England Nuclear) per ml. Procollagen and partially processed collagen from which the The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact. 3302NH2 propeptide was removed (...

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“…Alternatively, the pH 5.0 C-i product may result from cleavage at a different site than the position 6on7. Scott and Pearson (22) assayed only for cleavage occurring NH2-terminal to Lys i6C in the telopeptide. Our data with cleavage at pH <5.0, indicating that the released COOH-terminal portion is about 1.2 kDa larger than C-i suggests cleavage at l6C-i7c.…”

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Cathepsin D-mediated processing of procollagen: lysosomal enzyme involvement in secretory processing of procollagen.

Helseth¹,

Veis²

1984

Proc. Natl. Acad. Sci. U.S.A.

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“…We have previously reported [1] that the lysosomal proteinase cathepsin D catalyzes hydrolysis of the a1 chain of native or denatured type I collagen within the extra-helical carboxy-terminal sequence. It was found that double-chain peptides joined by cross-links in which the (6-hydroxy)a-amino-adipic-hemialdehyde residue at position C37 [2] is involved, were cleaved by treatment with enzyme.…”

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Cathepsin D: Specificity of Peptide‐Bond Cleavage in Type‐I Collagen and Effects on Type‐III Collagen and Procollagen

Scott

Pearson

1981

European Journal of Biochemistry

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Cathepsin D, purified from bovine thymus, has a limited proteolytic effect on types I and III bovine collagens. The α1 (I) chain was cleaved in native or denatured collagen only within the carboxy‐terminal extrahelical sequence, the major site being between residues C6 (Leu) and C7 (Ser). The α2 chain was unaffected in native collagen but was slowly cleaved between residues 782 (Phe) and 783 (Leu) in the denatured form. Cleavages, at 45° C, in type III collagen occur within the extra‐helical amino‐terminal sequence, on the carboxyterminal side of the lysine residue involved in intermolecular cross‐linking. All three sites of action are within sequences of general hydrophobic character. The very restricted cleavage of peptide bonds in denatured collagens can be ascribed to the infrequent occurrence of groupings of more than two hydrophobic residues and to the high content of the conformation limiting residues proline and hydroxyproline. The previously demonstrated failure of cathepsin D to solubilize a representative proportion of type III collagen from the fibres of bovine skin collagen (P. G. Scott and C. H. Pearson (1978) Biochem. Soc. Trans. 6, 1197–1199] may be explained by lack of ability of the enzyme to act on this collagen at 25° C, in such a manner as to separate molecules joined by intermolecular cross‐links involving the amino‐terminal extrahelical region of the molecule.

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“…Since the mechanical strength of venous walls depends to a large extent on the content of insoluble proteins such as elastin and collagen, and on their remaining intact, some authors consider that in the etiology of varicose veins degradation of the elastin in the venous walls by elastase is a factor leading to the development of varicosities [8]. As we know, elastin and collagen are degraded by cathepsin D [2,[12][13][14] and this enzyme occurs in the walls of normal veins [7], normal [6,7], and atheromatous arteries [ 1 ] and in venous grafts [4], There is, however, no data on the activity of cathepsin D in the walls of varicose veins.…”

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Cathepsin D Activity and Protein Degradation Products Content in the Walls of Varicose Veins of the Lower Limbs

Głowiński

Worowski

1981

Eur Surg Res

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Cathepsin D activity and the content of proteins and acid-soluble tyrosine in the walls of varicose veins, thrombophlebitic varicose veins and normal saphenous veins were determined. It was found that the cathepsin D activity in the wall of varicose veins is almost 1.5 times as high as that activity in a thrombophlebitic varicose vein wall and more than 3 times higher than in the wall of a normal vein. The acid-soluble tyrosine content of the varicose vein was slightly higher than that of the normal vein, whereas the content of the thrombophlebitic varicose vein was more than 1.5 times as high as that of the normal vein. Cathepsin D has a destructive effect on the varicose vein wall and accelerates the removal of thrombi in varicophlebitis.

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Cathepsin D: Cleavage of soluble collagen and crosslinked peptides

Cited by 25 publications

References 17 publications

Cathepsin D-mediated processing of procollagen: lysosomal enzyme involvement in secretory processing of procollagen.

Cathepsin D-mediated processing of procollagen: lysosomal enzyme involvement in secretory processing of procollagen.

Cathepsin D: Specificity of Peptide‐Bond Cleavage in Type‐I Collagen and Effects on Type‐III Collagen and Procollagen

Cathepsin D Activity and Protein Degradation Products Content in the Walls of Varicose Veins of the Lower Limbs

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