title: NMR solution structure of the catalytic fragment of human fibroblast collagenase complexed with a sulfonamide derivative of a hydroxamic acid compound.
abstract: 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 been determined using two-dimensional and three-dimensional heteronuclear NMR spectroscopy. The solution structure of the complex was calculated by means of hybrid distance geometry-simulated annealing using a 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 moiety of CGS-27023A was found to chelate to the "right" of the catalytic zinc where the p-methoxyphenyl sits in the S1' active-site pocket, the isopropyl group is in contact with H83 and N80, and the pyridine ring is solvent exposed. The sulfonyl oxygens are in hydrogen-bonding distance to the backbone NHs of L81 and 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 intramolecular restraints were observed for CGS-27023A from a 2D 12C-filtered NOESY experiment. A minimal set of NMR experiments in combination with the free MMP-1 assignments were used to assign the MMP-1 (1)H, 13C, and 15N resonances in the MMP-1:CGS-27023A complex. The assignments of CGS-27023A in the complex were obtained from 2D 12C-filtered NOESY and 2D 12C-filtered TOCSY experiments.
authors.affiliation: Department of Structural Biology, Wyeth-Ayerst Research, Pearl River, New York 10965, USA.
authors.forename: F J
authors.initials: FJ
authors.lastname: Moy
authors.forename: P K
authors.initials: PK
authors.lastname: Chanda
authors.forename: J M
authors.initials: JM
authors.lastname: Chen
authors.forename: S
authors.initials: S
authors.lastname: Cosmi
authors.forename: W
authors.initials: W
authors.lastname: Edris
authors.forename: J S
authors.initials: JS
authors.lastname: Skotnicki
authors.forename: J
authors.initials: J
authors.lastname: Wilhelm
authors.forename: R
authors.initials: R
authors.lastname: Powers
mesh_terms: D002384:Catalysis; D017364:Collagenases / Q000737:chemistry*; D003198:Computer Simulation; D018360:Crystallography, X-Ray; D005347:Fibroblasts / Q000201:enzymology*; D006801:Humans; D006877:Hydroxamic Acids / Q000737:chemistry*; D046911:Macromolecular Substances; D020781:Matrix Metalloproteinase 1; D019278:Matrix Metalloproteinase 3 / Q000737:chemistry; D020784:Matrix Metalloproteinase 8; D061965:Matrix Metalloproteinase Inhibitors; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D010446:Peptide Fragments / Q000037:antagonists & inhibitors / Q000737:chemistry*; D011480:Protease Inhibitors / Q000737:chemistry*; D011487:Protein Conformation; D011719:Pyrazines; D012996:Solutions; D013449:Sulfonamides / Q000737:chemistry*
chemical_list: C097658:CGS 27023A; D006877:Hydroxamic Acids; D046911:Macromolecular Substances; D061965:Matrix Metalloproteinase Inhibitors; D010446:Peptide Fragments; D011480:Protease Inhibitors; D011719:Pyrazines; D012996:Solutions; D013449:Sulfonamides; D017364:Collagenases; D019278:Matrix Metalloproteinase 3; D020784:Matrix Metalloproteinase 8; D020781:Matrix Metalloproteinase 1