caa a22 4500 47713579X CHVBK 20180405111717.0 cr unu---uuuuu 170330e19970701xx s 000 0 eng 10.1023/A:1018314808361 doi (NATIONALLICENCE)springer-10.1023/A:1018314808361 Alterations in chemical shifts and exchange broadening upon peptide boronic acid inhibitor binding to α-lytic protease [Elektronische Daten] [Jonathan Davis, David Agard, Tracy Handel, Vladimir Basus] α-Lytic protease, a bacterial serine protease of 198 aminoacids (19800 Da), has been used as a model system for studies of catalyticmechanism, structure-function relationships, and more recently forstudies of pro region-assisted protein folding. We have assigned thebackbones of the enzyme alone, and of its complex with the tetrahedraltransition state mimic N-tert-butyloxycarbonyl-Ala-Pro-boroVal, usingdouble- and triple-resonance 3D NMR spectroscopy on uniformly15N- and 13C/15N-labeled protein.Changes in backbone chemical shifts between the uncomplexed and inhibitedform of the protein are correlated with distance from the inhibitor, thedisplacement of backbone nitrogens, and change in hydrogen bond strengthupon inhibitor binding (derived from previously solved crystal structures).A comparison of the solution secondary structure of the uninhibited enzymewith that of the X-ray structure reveals no significant differences.Significant line broadening, indicating intermediate chemical exchange, wasobserved in many of the active site amides (including three broadened toinvisibility), and in a majority of cases the broadening was reversed uponaddition of the inhibitor. Implications and possible mechanisms of this linebroadening are discussed. Kluwer Academic Publishers, 1997 Serine protease nationallicence 3D NMR nationallicence Heteronuclear NMR nationallicence Davis Jonathan Graduate Group in Biophysics, University of California, 94143-0448, San Francisco, CA, U.S.A aut Agard David Graduate Group in Biophysics, University of California, 94143-0448, San Francisco, CA, U.S.A aut Handel Tracy Department of Molecular and Cellular Biology, University of California, 94720, Berkeley, CA, U.S.A aut Basus Vladimir Department of Pharmaceutical Chemistry, University of California, 94143-0446, San Francisco, CA, U.S.A aut Journal of Biomolecular NMR Kluwer Academic Publishers 10/1(1997-07-01), 21-27 0925-2738 10:1<21 1997 10 10858 https://doi.org/10.1023/A:1018314808361 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1018314808361 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Davis Jonathan Graduate Group in Biophysics, University of California, 94143-0448, San Francisco, CA, U.S.A aut NATIONALLICENCE 700 1- Agard David Graduate Group in Biophysics, University of California, 94143-0448, San Francisco, CA, U.S.A aut NATIONALLICENCE 700 1- Handel Tracy Department of Molecular and Cellular Biology, University of California, 94720, Berkeley, CA, U.S.A aut NATIONALLICENCE 700 1- Basus Vladimir Department of Pharmaceutical Chemistry, University of California, 94143-0446, San Francisco, CA, U.S.A aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Kluwer Academic Publishers 10/1(1997-07-01), 21-27 0925-2738 10:1<21 1997 10 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer