Effects of heparin and cholesterol sulfate on the activity and stability of human matrix metalloproteinase 7

Abstract

Abstract Sulfated glycosaminoglycans and sulfated lipids are involved in the biological functions of human matrix metalloproteinase 7 (MMP-7). In this study, the effects of heparin and cholesterol sulfate (CS) on the activity and stability of MMP-7 in the hydrolysis of a synthetic substrate, (7-methoxycoumarin-4-yl)acetyl-l-Pro-l-Leu-Gly-l-Leu-[N3-(2,4-dinitrophenyl)-l-2,3-diaminopropionyl]-l-Ala-l-Arg-NH2, were examined. Heparin increased activity by decreasing Km, and the Km values for 0 and 50 μM heparin were 57 ± 8 and 19 ± 5 μM, respectively. CS decreased activity in a non-competitive inhibitory manner with a Ki value of 11 ± 3 μM. In thermal incubation at 50−70 °C, heparin increased relative activity (the ratio of kcat/Km of MMP-7 with incubation to that without it), while CS decreased relative activity. These results indicate that heparin increases the activity and stability of MMP-7, while CS decreases them.