Andrew P. Hinck
University of Wisconsin-Madison
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Pure and Applied Chemistry | 1994
John L. Markley; Andrew P. Hinck; Stewart N. Loh; Kenneth E. Prehoda; Dagmar M. Truckses; William F. Walkenhorst; Jinfeng Wang
We have used NMR spectroscopy to determine peptide bond configurations and to measure the rates and equilibria of interconversion at individual Xaa-Pro peptide bond linkages in staphylococcal nuclease and several variants produced by site-directed mutagenesis. In general, tertiary interactions, rather than short-range interactions, have been found to be critical for stabilizing the cis linkage at Lys1I6- Pro1 which predominates in the dd-tge eye. A correlation has been found between the position of the cis/trans equilibrium at the Lys -Pro peptide bond and thermal stability of the variant. Enthalpic interactions that stabilize the folded protein appear to be present when the peptide bond is cis but not when it is trans. Hydrogen exchange protection factors correlate with the mole fraction of the protein that is in the cis configuration. Nuclease variants in which the peptide bond is predominantly cis are more stable against denaturation (by heat, pressure, or guanidinium chloride) than those that are predominantly truns. Disulfide bridges have been engineered and introduced by mutagenesis that stabilize certain conformational states; one of these shows coupling between the oxidation state of the engineered cysteine pair and the cislfruns configuration about the Ly~~-Pro~ peptide bond. A rough correlation is seen between the catalytic activities of mutants and the cisltrans ratio; the effect is primarily on &t rather than on Km.
Biochemistry | 1993
Catherine A. Royer; Andrew P. Hinck; Stewart N. Loh; Kenneth E. Prehoda; Xiangdong Peng; J. Jonas; John L. Markley
Journal of the American Chemical Society | 1992
Eric S. Eberhardt; Stewart N. Loh; Andrew P. Hinck; Ronald T. Raines
Biochemistry | 1990
Andrei T. Alexandrescu; Andrew P. Hinck; John L. Markley
Biochemistry | 1990
Jinfeng Wang; Andrew P. Hinck; Stewart N. Loh; John L. Markley
Biochemistry | 1992
Jinfeng Wang; Andrew P. Hinck; Stewart N. Loh; David M. LeMaster; John L. Markley
Biochemistry | 1996
Andrew P. Hinck; Dagmar M. Truckses; John L. Markley
Biochemistry | 1993
Andrew P. Hinck; Eric S. Eberhardt; John L. Markley
Biochemistry | 1990
Jinfeng Wang; Andrew P. Hinck; Stewart N. Loh; John L. Markley
Protein Engineering | 1993
Andrew P. Hinck; William F. Walkenhorst; William M. Westler; Soheui Choe; John L. Markley