Mechanistic aspects of cyanogenesis from active-site mutant Ser80Ala of hydroxynitrile lyase from Manihot esculenta in complex with acetone cyanohydrin

Protein Science : a Publication of the Protein Society
Hanspeter LaubleFranz Effenberger

Abstract

The structure and function of hydroxynitrile lyase from Manihot esculenta (MeHNL) have been analyzed by X-ray crystallography and site-directed mutagenesis. The crystal structure of the MeHNL-S80A mutant enzyme has been refined to an R-factor of 18.0% against diffraction data to 2.1-A resolution. The three-dimensional structure of the MeHNL-S80A-acetone cyanohydrin complex was determined at 2.2-A resolution and refined to an R-factor of 18.7%. Thr11 and Cys81 involved in substrate binding have been substituted by Ala in site-directed mutagenesis. The kinetic measurements of these mutant enzymes are presented. Combined with structural data, the results support a mechanism for cyanogenesis in which His236 as a general base abstracts a proton from Ser80, thereby allowing proton transfer from the hydroxyl group of acetone cyanohydrin to Ser80. The His236 imidazolium cation then facilitates the leaving of the nitrile group by proton donating.

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·F SangerA R Coulson
Oct 1, 1987·Analytical Biochemistry·D SelmarE E Conn
Mar 25, 1999·Acta Crystallographica. Section D, Biological Crystallography·H LaubleF Effenberger
Nov 5, 1999·Protein Science : a Publication of the Protein Society·J ZueggC Kratky
Sep 1, 1994·Acta Crystallographica. Section D, Biological Crystallography·UNKNOWN Collaborative Computational Project, Number 4

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Citations

Dec 10, 2003·Current Opinion in Biotechnology·Michael Breuer, Bernhard Hauer
Mar 9, 2004·Phytochemistry·Carsten Milkowski, Dieter Strack
Dec 14, 2001·Protein Science : a Publication of the Protein Society·Hanspeter LaubleFranz Effenberger
Jan 16, 2002·Protein Science : a Publication of the Protein Society·Ingrid DrevenyKarl Gruber
Apr 30, 2011·Annual Review of Plant Biology·Richard SayrePeng Zhang
Apr 7, 2005·Chembiochem : a European Journal of Chemical Biology·Holger BühlerFranz Effenberger
Mar 5, 2003·Chembiochem : a European Journal of Chemical Biology·Holger BühlerHarald Wajant
Aug 22, 2012·Natural Product Reports·Santosh PanjikarHeribert Warzecha

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