Cooperative binding of calcium ions modulates the tertiary structure and catalytic activity of Matrix-Metalloproteinase-9
Cooperative binding of calcium ions modulates the tertiary structure and catalytic activity of Matrix-Metalloproteinase-9作者机构：Department of Chemistry and Biochemistry North Dakota State University Fargo USA
出 版 物：《Advances in Enzyme Research》 (酶研究进展（英文）)
年 卷 期：2013年第1卷第2期
页 面：17-29页
学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学]
主 题：Matrix Metalloproteinase-9 Gelatinase-B Fluorescence Lifetime Quenching Acrylamide Stern-Volmer Constant
摘 要：To ascertain the molecular basis of Ca2+-mediated activation of matrix metalloproteinase-9 (MMP-9), we determined the accessibility of tryptophan residues to externally added acrylamide as quencher in the absence and presence of the metal ion. The steady-state and time resolved fluorescence data revealed that MMP-9 possesses two classes of tryptophan residues, “exposed and “buried which are quenched by the collisional rate constants (kq) of 3.2′ 109M-1.s-1 and 7.5′ 108M-1.s-1, respectively. These values are impaired by approximately two and three-fold, respectively, in the presence of 10 mM Ca2+. The Stern-Volmer constants (Ksv values) predicted from the time resolved fluorescence data (in the absence of Ca2+ ) satisfied the dynamic quenching model of the enzyme’s tryptophan residues. This was not the case in the presence of Ca2+;the steady-state acrylamide quenching data could only be explained by a combination of “dynamic and “static quenching models. A cumulative account of these data led to the suggestion that the binding of Ca2+ modulated the tertiary structure of the protein by decreasing the dynamic flexibility of the enzyme, which is manifested in further structuring of the enzyme’s active site pocket toward facilitating catalysis. Arguments are presented that the binding of Ca2+ at distal sites “dynamically communicates with the active site residues of MMP-9 during catalysis.
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