The proteolytic activity of the recombinant cryptic human fibronectin type IV collagenase from E-coli expression

Schnepel J, Tschesche H (2000)
JOURNAL OF PROTEIN CHEMISTRY 19(8): 685-692.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Abstract / Bemerkung
Human plasma ftbronectin (pFN) contains a cryptic metalloprotease present in the collagen-binding domain. The enzyme could be generated and activated in the presence of Ca2+ from the purified 70-kDa pFN fragment produced by cathepsin D digestion. In this work we cloned and expressed the metalloprotease, designated FN type IV collagenase (FnColA), and a truncated variant (FnColB) in E. coli. The recombinant pFN protein fragment was isolated from inclusion bodies, and subjected to folding and autocatalytic degradation in the presence of Ca2+, and yielded an active enzyme capable of digesting gelatin, helical type II and type IV collagen, alpha- and beta -casein, insulin b-chain, and a synthetic Mca-peptide. In contrast, isolated plasma fibronectin, type I collagen, and the DNP-peptide were no substrates. Both catalytically active recombinant pFN fragments were efficiently inhibited by EDTA, and batimastat, and, in contrast to the glycosylated enzyme isolated from plasma fibronectin, were also inhibited by TIMP-2.
Stichworte
type IV collagenase; TIMP-2; metalloprotease; fibronectin
Erscheinungsjahr
2000
Zeitschriftentitel
JOURNAL OF PROTEIN CHEMISTRY
Band
19
Ausgabe
8
Seite(n)
685-692
ISSN
0277-8033
Page URI
https://pub.uni-bielefeld.de/record/1617697

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Schnepel J, Tschesche H. The proteolytic activity of the recombinant cryptic human fibronectin type IV collagenase from E-coli expression. JOURNAL OF PROTEIN CHEMISTRY. 2000;19(8):685-692.
Schnepel, J., & Tschesche, H. (2000). The proteolytic activity of the recombinant cryptic human fibronectin type IV collagenase from E-coli expression. JOURNAL OF PROTEIN CHEMISTRY, 19(8), 685-692. doi:10.1023/A:1007104420017
Schnepel, J., and Tschesche, H. (2000). The proteolytic activity of the recombinant cryptic human fibronectin type IV collagenase from E-coli expression. JOURNAL OF PROTEIN CHEMISTRY 19, 685-692.
Schnepel, J., & Tschesche, H., 2000. The proteolytic activity of the recombinant cryptic human fibronectin type IV collagenase from E-coli expression. JOURNAL OF PROTEIN CHEMISTRY, 19(8), p 685-692.
J. Schnepel and H. Tschesche, “The proteolytic activity of the recombinant cryptic human fibronectin type IV collagenase from E-coli expression”, JOURNAL OF PROTEIN CHEMISTRY, vol. 19, 2000, pp. 685-692.
Schnepel, J., Tschesche, H.: The proteolytic activity of the recombinant cryptic human fibronectin type IV collagenase from E-coli expression. JOURNAL OF PROTEIN CHEMISTRY. 19, 685-692 (2000).
Schnepel, J, and Tschesche, Harald. “The proteolytic activity of the recombinant cryptic human fibronectin type IV collagenase from E-coli expression”. JOURNAL OF PROTEIN CHEMISTRY 19.8 (2000): 685-692.

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PMID: 11307953
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