PMID: 6161642Dec 9, 1980Paper

Carbon-13 nuclear magnetic resonance studies of the selectively isotope-labeled reactive site peptide bond of the basic pancreatic trypsin inhibitor in the complexes with trypsin, trypsinogen, and anhydrotrypsin

Biochemistry
R RicharzK Wüthrich

Abstract

A previously characterized modification of the basic pancreatic trypsin inhibitor (BPTI), with the carbonyl carbon atom of Lys-15 selectively enriched in 13C, the peptide bond Arg-39--Ala-40 cleaved, and Arg-39 removed, was used for 13C NMR studies of the reactive site peptide bond Lys-15--Ala-16 in the complexes with trypsin, trypsinogen, and anhydrotrypsin. The chemical shift of [1-13C]Lys-15 was 175.7 ppm in the free inhibitor, 176.4 ppm in the complexes with trypsin and anhydrotrypsin and the ternary complex with trypsinogen and H-Ile-Val-OH, and 175.7 ppm in a neutral solution containing the inhibitor and trypsinogen. These data show that the trypsin--BPTI complex does not contain a covalent tetrahedral carbon atom in the position of the reactive site peptide carbonyl of the inhibitor. They would be consistent with the formation of a noncovalent complex but cannot at present be used to further characterize the degree of a possible pyramidalization of the carbonyl carbon of Lys-15 in such a complex. The identical chemical shifts in the complexes with trypsin and anhydrotrypsin indicate that the gamma-hydroxyl group of Ser-195 of trypsin does not have an important role in the binding of the inhibitor. The previously describe...Continue Reading

References

Mar 15, 1979·Biochemical and Biophysical Research Communications·M W HunkapillerJ H Richards
Mar 1, 1976·Archives of Biochemistry and Biophysics·V Kasche
Mar 12, 1975·Helvetica Chimica Acta·C GrathwohlK Wüthrich
Sep 24, 1974·Biochemistry·J P VincentM Lazdunski
Nov 30, 1967·Biochemical and Biophysical Research Communications·T Chase, E Shaw

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Citations

May 1, 1990·Quarterly Reviews of Biophysics·D M LeMaster
Nov 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·C A McPhalenM N James
Dec 13, 2006·Journal of Molecular Biology·W Miachel HansonDavid P Goldenberg
Aug 12, 2008·Journal of Molecular Biology·Elena ZakharovaDavid P Goldenberg

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