Crystal versus solution structures of enzymes: NMR spectroscopy of a crystalline serine protease

Science
S O SmithWilliam W Bachovchin

Abstract

The hydrogen-bonding status of His57 in the catalytic triad (Asp-His-Ser) of serine protease has important mechanistic implications for this class of enzymes. Recent nitrogen-15 nuclear magnetic resonance (NMR) studies of alpha-lytic protease find His57 and Ser195 to be strongly hydrogen-bonded, a result that conflicts with the corresponding crystallographic studies, thereby suggesting that the crystal and solution structures may differ. This discrepancy is addressed and resolved in a nitrogen-15 NMR study of the enzyme in the crystalline state. The results show that the His-Ser and Asp-His interactions are identical in crystals and solutions, but that in crystals His57 titrates with a pKa of 7.9, nearly one pKa unit higher than in solution. This elevated pKa accounts for the absence of the His-Ser hydrogen bond in previous x-ray studies.

References

Nov 15, 1979·Journal of Molecular Biology·A R SieleckiM N James
Jan 1, 1977·Annual Review of Biochemistry·J Kraut
Apr 7, 1988·Nature·P Carter, J A Wells
Aug 28, 1969·Journal of Molecular Biology·G H CohenD R Davies
Jan 1, 1982·Annual Review of Biophysics and Bioengineering·T A Steitz, R G Shulman

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Citations

Jan 1, 1992·Proteins·P L HowellJ Hajdu
Aug 31, 2013·Journal of Biomolecular NMR·Marco FragaiEnrico Ravera
Mar 17, 2005·Journal of Molecular Biology·Ashish B PatelSteven O Smith
Dec 1, 1996·Quarterly Reviews of Biophysics·S O SmithM Groesbeek
Apr 18, 2012·Chemical Reviews·Awanish Kumar, Pannuru Venkatesu
Nov 19, 2011·Journal of the American Chemical Society·Anand BalakrishnanFrank Jordan
Dec 24, 2010·The Journal of Physical Chemistry. B·Sivakumar ParamasivamTatyana Polenova
Sep 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·S Farr-JonesW W Bachovchin
Apr 25, 2006·Proceedings of the National Academy of Sciences of the United States of America·Jun HuTimothy A Cross
Jul 9, 2013·Chemistry : a European Journal·Julien LeclaireGérard Buono
Aug 12, 2005·Chembiochem : a European Journal of Chemical Biology·Karsten SeidelMarc Baldus
Feb 12, 2005·Journal of Magnetic Resonance·John A StringerChad M Rienstra
Dec 1, 1990·Archives of Biochemistry and Biophysics·J SyguschM Allaire
Jan 7, 1998·Journal of Molecular Biology·S C MosimannM N James
Apr 20, 1993·Biochemistry·M AugerR G Griffin
Sep 26, 2002·Journal of the American Chemical Society·Bradley A KatzJames Janc
Apr 22, 2004·Journal of the American Chemical Society·Tatyana I IgumenovaAnn E McDermott
Jun 11, 2008·Journal of the American Chemical Society·Dirk NoltingBernd Winter
May 27, 2004·Journal of the American Chemical Society·Tatyana I IgumenovaA Joshua Wand

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