Abstract
II. Isolation of the metalloproteinase MPB. Comparison of the proteolytic activity on natural substrates by MPB, MSP 1 and MSP 2. Toxicon 31, 483–492, 1993.— A basic metalloproteinase active on casein was isolated from Bothrops moojeni venom by chromatography on Sephadex G-100, DEAE-Sephacel, SP-Sephadex C-50 and Sepharose 12. The enzyme, named MPB, is not hemorrhagic and presents only traces of blood-clotting activity. On polyacrylamide gel electrophoresis at pH 4.3, MPB presented a single and diffuse protein band. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the enzyme presented two protein bands corresponding to mol. wts of 65,000 and 55,000, which stained with Schiff's reagent. The proteolytic activity of MPB was inhibited by ethylenediaminetetracetate, 1,10-phenanthroline and dithiothreitol. The proteolytic activity of MPB and the serine proteinases MSP 1 and MSP 2 on natural substrates indicates differences in hydrolytic specificity among these enzymes. All fibrinogen chains were degraded by the three proteinases, but MPB is the most active. On fibrin, the proteinases hydrolyzed only the alpha-chain and alpha-polymer, leaving the beta-chain and gamma-dimer apparently untouched. The native type I collagen was partially hydrolyzed by the three enzymes but no digestion product was detected. On the contrary, calf and guinea-pig skin type I gelatins were readily digested by MSP 1 and MSP 2 producing different hydrolysis patterns. MPB was the least active proteinase on the gelatins. The digestion of fibronectin showed an inversion in the specificity of these proteinases. MPB was the most active on fibronectin, while MSP 1 and MSP 2 promoted a faint, partial hydrolysis on this protein.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.