Studies on Collagen

Seifter, Sam; Gallop, Paul M.; Klein, L.; Meilman, Edward

doi:10.1016/s0021-9258(18)70290-1

Cited by 116 publications

(9 citation statements)

References 16 publications

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“…Our data indicate that when the activities toward collagen and gelatin are compared in assays in which the same mass of substrate is present, gelatin is hydrolyzed 5-10 times faster than native collagens. This is in good agreement with the data of Seifter et al (1959). This difference in rates could be ascribed, in part, to the lower molarity of substrate (triple-helical rods) for native collagen compared to gelatin (mixture of component chains).…”

Section: Discussionsupporting

confidence: 91%

“…Over the years, many different molecular weights have been reported for Clostridial collagenases. Without specifying the designation given to the collagenase or the method used to determine the molecular weight, values that have been reported include 57 400 (Harper et al, 1965), 66000 (Keil, 1979), 70000 (Emod et al, 1981), 72000 (Lwebuga-Mukasa et al, 1976), 79000 (Yoshida & Noda, 1965), 81000 (Lwebuga et al, 1976), 95 000 (Yoshida & Noda, 1965), 105000 (Harper et al, 1965), 109 000 (Seifter et al, 1959;Strauch & Grassman, 1966), 112000 (Mandl et al, 1964), and 147000 (Soru & Zaharia, 1972). There is little doubt that some of the quoted values differ due to the fact that investigators have simply isolated different individual or subsets of the individual collagenases.…”

Section: Discussionmentioning

confidence: 99%

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Characterization of the individual collagenases from Clostridium histolyticum

Bond¹,

Wart²

1984

Biochemistry

212

141

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The six collagenases (alpha, beta, gamma, delta, epsilon, and zeta) from Clostridium histolyticum isolated in the preceding paper [Bond, M. D., & Van Wart, H. E. (1984) Biochemistry (first paper of three in this issue)] have been characterized in detail. The molecular weights determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis range from 68 000 to 125 000. Isoelectric focusing experiments demonstrate that the isoelectric points of the collagenases are in the 5.35-6.20 range. These experiments also reveal that the subspecies of alpha- and gamma-collagenases (alpha1 vs. alpha 2 and gamma 1 vs. gamma 2) have different isoelectric points but the same molecular weights. Microheterogeneity is also observed for the beta- and epsilon-collagenases. The amino acid compositions of all six collagenases have been determined, and analysis for neutral sugars and hexosamines shows that none of the enzymes have a significant carbohydrate content. Zinc and calcium are the only metals that copurify with the collagenases. The purified enzymes contain approximately 1 mol of zinc/mol of protein and a calcium content that varies from about 2 mol/mol for alpha-collagenase to about 7 mol/mol for beta-collagenase. All of the collagenases are 5-10 times more active against gelatin than collagen. The alpha-, beta-, and gamma-collagenases are significantly less active toward the synthetic peptide substrates examined than the delta-, epsilon, and zeta-collagenases. This property, taken together with data on the stabilities and amino acid compositions of these enzymes, strongly supports their assignment to two distinct classes. This establishes clearly that C. histolyticum does, indeed, produce more than one different type of collagenase.

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Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

Characterization of the individual collagenases from Clostridium histolyticum

Bond¹,

Wart²

1984

Biochemistry

212

141

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“…6 This collagen, dissolved in pYL 3.7 citrate buffer, and the parent gelatin produced from it by mild heating, have since been the object of extensive physico-chemical examination by Gallop,6•7 Cohen8 and especially Boedtker and Doty.9 From these studies it has been concluded that ichthyocol collagen exists in this solvent as rigid rod-like, three-stranded macromolecules (molecular weight approximately 4 X 106) and that these macromolecules dissociate on heating to form the singlechain, essentially randomly coiled molecules of parent gelatin. 10 Recently, a very active collagenase has been prepared and purified from Clostridium• histolyticum.u• 16 This enzyme seems to be highly specific for collagen and gelatin severing these proteins at amino acid sequences having the general formula -Pro.X.Gly.Pro.Y-, between X and Gly,16-19 while attacking no other natural substrate so far tested. Since collagenase is activated by ionic calcium15•20 and inactivated by low pH, acidic citrate buffer could not be used as a collagen solvent for studies with this enzyme.…”

mentioning

confidence: 99%

An Enzymatic Examination of the Structure of the Collagen Macromolecule¹

Hippel¹,

Gallop²,

Seifter³

et al. 1960

J. Am. Chem. Soc.

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“…Digestion of Immunological Precipitates by Collagenase.-Precipitates formed by pTyrGel and anti-pTyrGel rabbit serum were washed three times with cold 0.14 M NaCl and suspended in 0.05 M Tris buffer, pH 7.4, containing 0.005 M CaCl2. Collagenase (95 units/mg, a gift from Dr. P. M. Gallop) was added to the suspension in an amount calculated to be approximately 1 unit/mg pTyrGel (Seifter et al, 1959).…”

Section: Methodsmentioning

confidence: 99%

Isolation and Fragmentation of Antibodies to Polytyrosyl Gelatin^*

Sela¹

1964

Biochemistry

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Antibodies to polytyrosyl gelatin were isolated immunospecifically by the digestion of the antigen-antibody precipitate with collagenase. Pieces of the antigen which remained bound to the antibodies and blocked their activity were removed by passing the antibody preparation through a Sephadex column at acid pH. The antibodies thus isolated were 6.5 S y-globulin and precipitated with added fresh antigen. By using radioactively labeled antigen it was possible to follow the antibody-combining sites by means of the pieces of the antigen which remained bound to the antibody before the passage through Sephadex. These antigen pieces were associated with fragments I and II of the antibody after fragmentation with papain, while fragment III was devoid of such pieces. After reduction with mercaptoethanol in aqueous solution and the separation of the subunits A and B of the antibody no radioactive pieces of the antigen were found in fraction B. The protein in fraction A, on the other hand, was associated with antigen pieces. Several alternative explanations for the presence of the antigen pieces in this fraction are discussed.

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Studies on Collagen

Cited by 116 publications

References 16 publications

Characterization of the individual collagenases from Clostridium histolyticum

Characterization of the individual collagenases from Clostridium histolyticum

An Enzymatic Examination of the Structure of the Collagen Macromolecule¹

Isolation and Fragmentation of Antibodies to Polytyrosyl Gelatin^*

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Studies on Collagen

Cited by 116 publications

References 16 publications

Characterization of the individual collagenases from Clostridium histolyticum

Characterization of the individual collagenases from Clostridium histolyticum

An Enzymatic Examination of the Structure of the Collagen Macromolecule1

Isolation and Fragmentation of Antibodies to Polytyrosyl Gelatin*

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Product

Resources

About

An Enzymatic Examination of the Structure of the Collagen Macromolecule¹

Isolation and Fragmentation of Antibodies to Polytyrosyl Gelatin^*