Examining the role of glutamic acid 183 in chloroperoxidase catalysis

The Journal of Biological Chemistry
Xianwen YiLowell P. Hager

Abstract

Site-directed mutagenesis has been used to investigate the role of glutamic acid 183 in chloroperoxidase catalysis. Based on the x-ray crystallographic structure of chloroperoxidase, Glu-183 is postulated to function on distal side of the heme prosthetic group as an acid-base catalyst in facilitating the reaction between the peroxidase and hydrogen peroxide with the formation of Compound I. In contrast, the other members of the heme peroxidase family use a histidine residue in this role. Plasmids have now been constructed in which the codon for Glu-183 is replaced with a histidine codon. The mutant recombinant gene has been expressed in Aspergillus niger. An analysis of the produced mutant gene shows that the substitution of Glu-183 with a His residue is detrimental to the chlorination and dismutation activity of chloroperoxidase. The activity is reduced by 85 and 50% of wild type activity, respectively. However, quite unexpectedly, the epoxidation activity of the mutant enzyme is significantly enhanced approximately 2.5-fold. These results show that Glu-183 is important but not essential for the chlorination activity of chloroperoxidase. It is possible that the increased epoxidation of the mutant enzyme is based on an increase...Continue Reading

References

Nov 1, 1979·The Biochemical Journal·M D Corbett, B R Chipko
Sep 8, 1992·Biochemistry·J C Alvarez, P R Ortiz de Montellano
Jan 6, 1992·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·P F Hollenberg
May 22, 1969·Biochemical and Biophysical Research Communications·J A Thomas, Lowell P. Hager
Apr 1, 1981·Canadian Journal of Biochemistry·T AraisoH B Dunford
Sep 26, 1995·Biochemistry·Q ZongLowell P. Hager
Jun 1, 1995·Archives of Biochemistry and Biophysics·V P MillerP R Ortiz de Montellano
Nov 12, 1996·Proceedings of the National Academy of Sciences of the United States of America·P G DebrunnerLowell P. Hager
Jan 8, 1999·Biochemical and Biophysical Research Communications·S Hu, Lowell P. Hager
Oct 27, 1999·Proceedings of the National Academy of Sciences of the United States of America·X YiLowell P. Hager
Jul 6, 2000·Applied and Environmental Microbiology·A ConesaP J Punt
May 31, 1996·The Journal of Organic Chemistry·Frederick J. Lakner, Lowell P. Hager
Sep 1, 1995·Acta Crystallographica. Section D, Biological Crystallography·M SundaramoorthyT L Poulos

Citations

Oct 2, 2012·The Journal of Physical Chemistry. B·Alexander N Morozov, David C Chatfield
May 22, 2007·BMC Biotechnology·Jian-Zhong Liu, Min Wang
Dec 24, 2013·Applied Biochemistry and Biotechnology·Haiyun LiQuanguo Zhai
May 10, 2003·Chemosphere·Emma JohanssonGunilla Oberg
May 4, 2017·Chemical Reviews·Jonathan LathamJason Micklefield
Mar 2, 2016·Archives of Biochemistry and Biophysics·Rui ZhangXiaotang Wang
Mar 11, 2010·The Journal of Biological Chemistry·Lowell P. Hager
Nov 9, 2005·Biotechnology and Bioengineering·Jin-Byung Park, Douglas S Clark
Jun 23, 2006·The Journal of Biological Chemistry·Karin KühnelIlme Schlichting

Related Concepts

Epoxidase
Aspergillus niger
Catalase T
Catalysis
Chloride Peroxidase
Chlorine
Ion-Exchange Chromatography Procedure
Circular Dichroism, Vibrational
SDS-PAGE
Filamentous fungus

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.