Immunopurification and characterization of a collagenase/gelatinase domain issued from basement membrane fibronectin

FEBS Letters

Boudjennah, Laziz¹  Ylätupa, Sari²  Dalet-Fumeron, Véronique¹  Pagano, Maurice¹ 

[1] Biochimie des protéases, Faculté de Médecine Broussais Hôtel-Dieu,Université Pierre et Marie Curie, 15 rue de l'école de médecine, 75270 Paris Cedex 06, France;Biohit Oy, Vienankatu 5, 87100 Kajaani, Finland

关键词: Cellular fibronectin;    Gelatinase;    Immunopurification;    Monoclonal antibody;    BSA;    bovine serum albumin;    DTE;    dithioerythritol;    CNBr;    cyanogen bromide;    MMP;    matrix metalloproteinase;    Dpa;    Mca;    (7-methoxycoumarin-4-yl)acetyl;    EDTA;    ethylene diamine tetraacetate;    disodium salt;   

DOI  :  10.1016/0014-5793(96)00699-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The proteolytic potential of cellular fibronectin fragments issued from a basement membrane hydrolysate was investigated. Three different gelatinase activities (47, 43 and 37 kDa), located by gelatin zymography, were isolated using successively heparin-agarose, gelatin-agarose and immunopurification with polyclonal antibodies directed against bovine plasma fibronectin. These fragments were also characterized using a monoclonal antibody directed against the extra-domain EDA of cellular fibronectin as a probe. A collagenase activity, reliably indicated by the gelatin zymography pattern, was also found using MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH₂, the intramolecularly quenched fluorogenic substrate of collagenases. From these results, cellular fibronectin was found to be able to exhibit a proteolytic function after limited proteolysis. This MMP-like function could be associated with tissue remodeling in both normal and pathological states, such as metastasis, angiogenesis and tissue repair.

【 授权许可】

Unknown