Studies on the Structure of Collagen Utilizing a Collagenolytic Enzyme from Tadpole

Andrew Kang, Yutaka Nagai, Karl A. Piez, Jerome Gross

Research output: Contribution to journalArticle

103 Citations (Scopus)

Abstract

Native rat skin collagen was digested with an enzyme from tadpole which is known to cleave the molecule into two unequal pieces, each of which retains the triple helical collagen structure. The larger piece (TO) represents 75 % and the smaller piece (TCB) 25 % of the length of the molecule. Denaturation of the digestion products and fractionation by molecular sieve and carboxymethylcellulose chromatography permitted the isolation of fragments which were shown by molecular weight determination and amino acid analysis to be derived from the cleavage of the two α1 and one α2 chains each into two pieces, representing one quarter and three quarters of the original chain. The small pieces, α1B and α2B, have molecular weights of 24,000. The large pieces, α1A and α2A, have molecular weights of 71,000. These results demonstrate that in cleaving the collagen molecule the enzyme cleaves all three α chains in the same proportion. Therefore, the three α chains must each extend the full length of the molecule. The presence in the denatured digestion mixture of a fraction with a molecular weight twice that of the large pieces and the absence of a fraction which could be a dimer of the small pieces indicate that interchain crosslinks occur only in TCA. Since an intramolecular cross-link has been shown to be near the N-terminal ends of the α chains, these, together with the cross-link, must be at the A end of the molecule. Analysis showed that many of the amino acid residues are unequally distributed between the A and B pieces.

Original languageEnglish (US)
Pages (from-to)509-515
Number of pages7
JournalBiochemistry
Volume5
Issue number2
DOIs
StatePublished - Jan 1 1966
Externally publishedYes

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Larva
Collagen
Molecular Weight
Molecules
Molecular weight
Enzymes
Digestion
Amino Acids
Carboxymethylcellulose Sodium
Denaturation
Gel Chromatography
Molecular sieves
Fractionation
Chromatography
Dimers
Rats
Skin

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Studies on the Structure of Collagen Utilizing a Collagenolytic Enzyme from Tadpole. / Kang, Andrew; Nagai, Yutaka; Piez, Karl A.; Gross, Jerome.

In: Biochemistry, Vol. 5, No. 2, 01.01.1966, p. 509-515.

Research output: Contribution to journalArticle

Kang, Andrew ; Nagai, Yutaka ; Piez, Karl A. ; Gross, Jerome. / Studies on the Structure of Collagen Utilizing a Collagenolytic Enzyme from Tadpole. In: Biochemistry. 1966 ; Vol. 5, No. 2. pp. 509-515.
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