Gelatinase activity in Mycobacterium bovis protein extract.
Proteases are well-recognized as virulence factors in different pathologies, resulting in tissue damage potential. Despite efforts over the past few years to identify mycobacterial protein antigens, there is little information regarding the role of mycobacterial proteinase activities. In this study, by zymography techniques, we have detected and partially studied some biochemical properties of Mycobacterium bovis proteases, such as pH dependency of activity and susceptibility to classical proteinase inhibitors. We observed optimal proteolytic activity at pH 8. Some proteinases were inhibited by classic inhibitors of serine proteases, such as PMSF, AEBSF, and 3-4 DCI. In some AEBSF pre-treated preparations we observed residual gelatinase activity in Rf 0.32. This gelatinase was stimulated by Zn2+ and inhibited by OPA (1 mM). This last effect was reversed by exposure to equimolar quantitative OPA/Zn+2 (1 mM/1 mM). These results suggest the existence of serine proteinase and metalloproteinase types in protein extracts of Mycobacterium bovis.
Rada
Elsa
E
Laboratorio de Leprología y Patología Experimental, Instituto de Biomedicina, Apartado 4043, Caracas 1010A, Venezuela. erada@telcel.net.ve
Zambrano
Edgar Armando
EA
Rodríguez
Ingrid Yocelin
IY
Santaella
Carlos
C
Convit
Jacinto
J
D000818:Animals; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002417:Cattle; D003600:Cytosol / Q000201:enzymology; D018093:Gelatinases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008565:Membrane Proteins / Q000378:metabolism; D009163:Mycobacterium bovis / Q000201:enzymology* / Q000254:growth & development; D011480:Protease Inhibitors / Q000494:pharmacology
D001426:Bacterial Proteins; D008565:Membrane Proteins; D011480:Protease Inhibitors; D018093:Gelatinases