Members of the cystatin superfamily interact with MMP-9 and protect it from autolytic degradation without affecting its gelatinolytic activities.
Matrix metalloproteinases (MMPs), like other proteinases, can undergo autolytic degradation once activated in vivo. Whereas the activities of these enzymes are tightly regulated by tissue inhibitors of matrix metalloproteinases (TIMPs), it is not clear mechanistically how these enzymes are protected from autolysis in their active state. We previously reported that MMPs particularly MMP-9 and MMP-2 interact with the serum glycoprotein fetuin-A [Arch. Biochem. Biophys. (1995) 322, 250], a member of the cystatin superfamily. In the present analyses, we demonstrate that this interaction protects MMP-9 from autolytic degradation without interfering with its enzymatic activity, allowing it to efficiently digest gelatin. Our data demonstrate that MMP-9 binds to members of the cystatin family with K(diss) ranging from 25 to 58 nM for fetuin-A and 1.5-1.9 microM for cystatin C. The ability of fetuin-A to protect MMP-9 from autolysis requires a molar ratio of at least 8:1 (fetuin-A/MMP-9). More interestingly, our data show that the other members of the cystatin also have the ability to protect MMP-9 from autolysis, provided they are in molar excess relative to MMP-9. Taken together, our data suggest that cystatins, particularly fetuin-A, in any cellular compartment including the circulatory system, efficiently protect MMP-9 and possibly other MMPs from autolysis. This mechanism ensures the digestion of the preferred substrate for MMP-9 without sacrificing the enzyme in the process.
Ray
Sanhita
S
Department of Biochemistry, Meharry Medical College, 1005 Dr. D.B. Todd Jr. Blvd., Nashville, TN 37208, USA.
Lukyanov
Pavel
P
Ochieng
Josiah
J
D000818:Animals; D001798:Blood Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D002417:Cattle; D055316:Cystatin C; D015891:Cystatins / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000378:metabolism; D006801:Humans; D007704:Kininogens / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D005810:Multigene Family; D060749:alpha-2-HS-Glycoprotein
C556354:AHSG protein, human; C556357:Ahsg protein, mouse; D001798:Blood Proteins; C525030:CST3 protein, human; C525031:Cst3 protein, mouse; D055316:Cystatin C; D015891:Cystatins; D015853:Cysteine Proteinase Inhibitors; D007704:Kininogens; D060749:alpha-2-HS-Glycoprotein; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9