caa a22 4500 475744403 CHVBK 20180406123509.0 cr unu---uuuuu 170329e20001101xx s 000 0 eng 10.1023/A:1007104420017 doi (NATIONALLICENCE)springer-10.1023/A:1007104420017 The Proteolytic Activity of the Recombinant Cryptic Human Fibronectin Type IV Collagenase from E. coli Expression [Elektronische Daten] [Jörn Schnepel, Harald Tschesche] Human plasma fibronectin (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, α- and β-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. Plenum Publishing Corporation, 2000 Fibronectin nationallicence type IV collagenase nationallicence metalloprotease nationallicence TIMP-2 nationallicence Schnepel Jörn Department of Biochemistry I, Faculty of Chemistry, University of Bielefeld, Germany aut Tschesche Harald Department of Biochemistry I, Faculty of Chemistry, University of Bielefeld, Germany aut Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/8(2000-11-01), 685-692 0277-8033 19:8<685 2000 19 10930 https://doi.org/10.1023/A:1007104420017 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1007104420017 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Schnepel Jörn Department of Biochemistry I, Faculty of Chemistry, University of Bielefeld, Germany aut NATIONALLICENCE 700 1- Tschesche Harald Department of Biochemistry I, Faculty of Chemistry, University of Bielefeld, Germany aut NATIONALLICENCE 773 0- Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/8(2000-11-01), 685-692 0277-8033 19:8<685 2000 19 10930 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer