NMR Solution Structure of the Catalytic Fragment of Human Fibroblast Collagenase Complexed with a Sulfonamide Derivative of a Hydroxamic Acid Compound

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• 作者：Franklin J. Moy ; Pranab K. Chanda ; James M. Chen ; Scott Cosmi ; Wade Edris ; Jerauld S. Skotnicki ; Jim Wilhelm ; and Robert Powers
• 刊名：Biochemistry
• 出版年：1999
• 出版时间：June 1, 1999
• 年：1999
• 卷：38
• 期：22
• 页码：7085 - 7096
• 全文大小：165K
• 年卷期：v.38,no.22(June 1, 1999)
• ISSN：1520-4995

文摘

The solution structure of the catalytic fragment of human fibroblast collagenase (MMP-1)complexed with a sulfonamide derivative of a hydroxamic acid compound (CGS-27023A) has beendetermined using two-dimensional and three-dimensional heteronuclear NMR spectroscopy. The solutionstructure of the complex was calculated by means of hybrid distance geometry-simulated annealing usinga combination of experimental NMR restraints obtained from the previous refinement of the inhibitor-free MMP-1 (1) and recent restraints for the MMP-1:CGS-27023A complex. The hydroxamic acid moietyof CGS-27023A was found to chelate to the "right" of the catalytic zinc where the p-methoxyphenyl sitsin the S1' active-site pocket, the isopropyl group is in contact with H83 and N80, and the pyridine ringis solvent exposed. The sulfonyl oxygens are in hydrogen-bonding distance to the backbone NHs of L81and A82. This is similar to the conformation determined by NMR of the inhibitor bound to stromelysin(2, 3). A total of 48 distance restraints were observed between MMP-1 and CGS-27023A from 3D ¹³C-edited/¹²C-filtered NOESY and 3D ¹⁵N-edited NOESY experiments. An additional 18 intramolecularrestraints were observed for CGS-27023A from a 2D ¹²C-filtered NOESY experiment. A minimal set ofNMR experiments in combination with the free MMP-1 assignments were used to assign the MMP-1 ¹H,¹³C, and ¹⁵N resonances in the MMP-1:CGS-27023A complex. The assignments of CGS-27023A in thecomplex were obtained from 2D ¹²C-filtered NOESY and 2D ¹²C-filtered TOCSY experiments.