Recombinant Cryptic Human Fibronectinase Cleaves Actin and Myosin: Substrate Specificity and Possible Role in Muscular Dystrophy
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Jörn Schnepel
Abstract
The Nterminal heparin/fibrin binding domain of human plasma fibronectin (pFN) contains a cryptic proteinase. The enzyme could be generated and activated in the presence of Ca 2+ from the purified 70 kDa pFN fragment produced by cathepsin D digestion of pFN. In this work we cloned and expressed the serine proteinase, designated fibronectinase (Fnase), in E. coli. The recombinant pFN protein fragment was isolated from inclusion bodies, subjected to folding and autocatalytic degradation in the presence of Ca 2+ , and yielded an active enzyme capable of digesting fibronectin. Cleavage of pFN and the synthetic peptides AcIEGKpNA and BzIEGRpNA demonstrated identical specificity of the recombinant and the isolated fibronectinase. Further investigations of the substrate specificity revealed for the first time the muscle proteins actin and myosin as being substrates of fibronectinase. The enzyme can be inhibited by α1-proteinase inhibitor. In the context of induced cathepsin D release, e. g. from granulocytes under inflammatory conditions, these results indicate an increase in specific proteolytic potential against muscular proteins in dystrophic diseases by the release of cryptic fibronectinase.
Copyright © 2001 by Walter de Gruyter GmbH & Co. KG
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- Erratum
- Acknowledgement
- Content Index
- Author Index
- Subject Index
Articles in the same Issue
- New Control of Mitochondrial Membrane Potential and ROS Formation A Hypothesis
- Brix from Xenopus laevis and Brx1p From Yeast Define a New Family of Proteins Involved in the Biogenesis of Large Ribosomal Subunits
- Mig-6 Is a Negative Regulator of the Epidermal Growth Factor Receptor Signal
- β-Carotene Inhibits Growth of Human Colon Carcinoma Cells in Vitro by Induction of Apoptosis
- Ligand-Mediated Protection against Phage Lysis as a Positive Selection Strategy for the Enrichment of Epitopes Displayed on the Surface of E. coli Cells
- Structural and Redox Properties of the Leaderless DsbE (CcmG) Protein: Both Active-Site Cysteines of the Reduced Form Are Involved in Its Function in the Escherichia coli Periplasm
- Polyphenols of Cocoa: Inhibition of Mammalian 15-Lipoxygenase
- Total Antioxidant Capacity and Nuclear DNA Damage in Keratinocytes after Exposure to H2 O2
- Recombinant Cryptic Human Fibronectinase Cleaves Actin and Myosin: Substrate Specificity and Possible Role in Muscular Dystrophy
- Rat Tripeptidyl Peptidase I: Molecular Cloning, Functional Expression, Tissue Localization and Enzymatic Characterization
- Determination of NADH in Frozen Rat Brain Sections by Laser-Induced Fluorescence
- Human -Calpain: Simple Isolation from Erythrocytes and Characterization of Autolysis Fragments
- Erratum
- Acknowledgement
- Content Index
- Author Index
- Subject Index
