caa a22 4500 477136273 CHVBK 20180405111719.0 cr unu---uuuuu 170330e19970101xx s 000 0 eng 10.1023/A:1018615400129 doi (NATIONALLICENCE)springer-10.1023/A:1018615400129 Assignments and structure determination of the catalytic domain of human fibroblast collagenase using 3D double and triple resonance NMR spectroscopy [Elektronische Daten] [Mark McCoy, Martin Dellwo, Diane Schneider, Tracey Banks, Joseph Falvo, Karen Vavra, Alan Mathiowetz, M. Qoronfleh, Richard Ciccarelli, Ewell Cook, Tricia Pulvino, Robert Wahl, Hsin Wang] We report here the backbone 1HN, 15N, 13Cα, 13CO, and 1Hα NMR assignmentsfor the catalytic domain of human fibroblast collagenase (HFC). Three independentassignment pathways (matching 1H, 13Cα, and 13CO resonances) were used to establishsequential connections. The connections using 13Cα resonances were obtained fromHNCOCA and HNCA experiments; 13CO connections were obtained from HNCO andHNCACO experiments. The sequential proton assignment pathway was established from a 3D(1H/15N) NOESY-HSQC experiment. Amino acid typing was accomplished using 13C and15N chemical shifts, specific labeling of 15N-Leu, and spin pattern recognition from DQF-COSY. The secondary structure was determined by analyzing the 3D (1H/15N) NOESY-HSQC. A preliminary NMR structure calculation of HFC was found to be in agreement withrecent X-ray structures of human fibroblast collagenase and human neutrophil collagenase aswell as similar to recent NMR structures of a highly homologous protein, stromelysin. Allthree helices were located; a five-stranded β-sheet (four parallel strands, one antiparallelstrand) was also determined. β-Sheet regions were identified by cross-stranddαN and dNN connections and by strong intraresidue dαN correlations, and were corroborated byobserving slow amide proton exchange. Chemical shift changes in a selectively 15N-labeledsample suggest that substantial structural changes occur in the active site cleft on the bindingof an inhibitor. Kluwer Academic Publishers, 1997 Collagenase nationallicence Assignments nationallicence Structure nationallicence MMP nationallicence 3D NMR nationallicence Matrix metalloproteinase nationallicence McCoy Mark Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut Dellwo Martin Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut Schneider Diane Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut Banks Tracey Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut Falvo Joseph Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut Vavra Karen Eastman Kodak Company, 14650, Rochester, NY, U.S.A aut Mathiowetz Alan Eastman Kodak Company, 14650, Rochester, NY, U.S.A aut Qoronfleh M. Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut Ciccarelli Richard Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut Cook Ewell Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut Pulvino Tricia Eastman Kodak Company, 14650, Rochester, NY, U.S.A aut Wahl Robert Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut Wang Hsin Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut Journal of Biomolecular NMR Kluwer Academic Publishers 9/1(1997-01-01), 11-24 0925-2738 9:1<11 1997 9 10858 https://doi.org/10.1023/A:1018615400129 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1018615400129 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- McCoy Mark Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut NATIONALLICENCE 700 1- Dellwo Martin Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut NATIONALLICENCE 700 1- Schneider Diane Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut NATIONALLICENCE 700 1- Banks Tracey Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut NATIONALLICENCE 700 1- Falvo Joseph Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut NATIONALLICENCE 700 1- Vavra Karen Eastman Kodak Company, 14650, Rochester, NY, U.S.A aut NATIONALLICENCE 700 1- Mathiowetz Alan Eastman Kodak Company, 14650, Rochester, NY, U.S.A aut NATIONALLICENCE 700 1- Qoronfleh M. Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut NATIONALLICENCE 700 1- Ciccarelli Richard Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut NATIONALLICENCE 700 1- Cook Ewell Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut NATIONALLICENCE 700 1- Pulvino Tricia Eastman Kodak Company, 14650, Rochester, NY, U.S.A aut NATIONALLICENCE 700 1- Wahl Robert Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut NATIONALLICENCE 700 1- Wang Hsin Sterling Winthrop Pharmaceuticals Research Division, 1250 South Collegeville Road, 19426, Collegeville, PA, U.S.A aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Kluwer Academic Publishers 9/1(1997-01-01), 11-24 0925-2738 9:1<11 1997 9 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer