Brassica juncea 3-hydroxy-3-methylglutaryl (HMG)-CoA synthase 1: expression and characterization of recombinant wild-type and mutant enzymes

The Biochemical Journal
Dinesh A NagegowdaMee-Len Chye

Abstract

3-hydroxy-3-methylglutaryl (HMG)-CoA synthase (HMGS; EC 2.3.3.10) is the second enzyme in the cytoplasmic mevalonate pathway of isoprenoid biosynthesis, and catalyses the condensation of acetyl-CoA with acetoacetyl-CoA (AcAc-CoA) to yield S-HMG-CoA. In this study, we have first characterized in detail a plant HMGS, Brassica juncea HMGS1 (BjHMGS1), as a His6-tagged protein from Escherichia coli. Native gel electrophoresis analysis showed that the enzyme behaves as a homodimer with a calculated mass of 105.8 kDa. It is activated by 5 mM dithioerythreitol and is inhibited by F-244 which is specific for HMGS enzymes. It has a pH optimum of 8.5 and a temperature optimum of 35 degrees C, with an energy of activation of 62.5 J x mol(-1). Unlike cytosolic HMGS from chicken and cockroach, cations like Mg2+, Mn2+, Zn2+ and Co2+ did not stimulate His6-BjHMGS1 activity in vitro; instead all except Mg2+ were inhibitory. His6-BjHMGS1 has an apparent K(m-acetyl-CoA) of 43 microM and a V(max) of 0.47 micromol x mg(-1) x min(-1), and was inhibited by one of the substrates (AcAc-CoA) and by both products (HMG-CoA and HS-CoA). Site-directed mutagenesis of conserved amino acid residues in BjHMGS1 revealed that substitutions R157A, H188N and C212S ...Continue Reading

References

Aug 1, 1991·Lipids·T J BachA Wettstein
Dec 1, 1990·Archives of Biochemistry and Biophysics·P A Kattar-CooleyH M Miziorko
Nov 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·M D GreenspanY C Chiang
Dec 14, 1987·Biochimica Et Biophysica Acta·H TomodaS Omura
Jan 1, 1967·Pure and Applied Chemistry. Chimie Pure Et Appliquée·F Lynen
Jun 1, 1997·Insect Biochemistry and Molecular Biology·J CabañóP F Marrero
Jun 10, 2000·The Plant Journal : for Cell and Molecular Biology·D AlexM L Chye
Sep 12, 2002·Annual Review of Plant Biology·André Kessler, Ian T Baldwin
Oct 24, 2002·The Journal of Biological Chemistry·Clare V SmithKerstin Höner zu Bentrup
Apr 22, 2003·Annual Review of Biochemistry·Daniel A KrautDaniel Herschlag
May 9, 2003·Natural Product Reports·Tomohisa Kuzuyama, Haruo Seto
May 14, 2003·The Journal of Biological Chemistry·Ila MisraHenry M Miziorko
May 16, 2003·Proceedings of the National Academy of Sciences of the United States of America·Oliver LauleMarkus Lange
Feb 27, 2004·Biochimica Et Biophysica Acta·Hiroshi TomodaSatoshi Omura
Mar 12, 2004·Annual Review of Plant Physiology and Plant Molecular Biology·Hartmut K. Lichtenthaler
Oct 21, 2004·The Plant Journal : for Cell and Molecular Biology·A Hemmerlin, T J Bach

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Citations

Mar 17, 2005·Planta·Dinesh A NagegowdaMee-Len Chye
Feb 11, 2010·BMC Bioinformatics·Aliah Hazmah Hawari, Zeti-Azura Mohamed-Hussein
Jul 26, 2006·Proceedings of the National Academy of Sciences of the United States of America·Florence PojerJoseph P Noel
Sep 23, 2016·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Ming GaoYangdong Wang
Apr 8, 2018·Applied and Environmental Microbiology·Christopher M Flynn, Claudia Schmidt-Dannert
Feb 4, 2012·The Arabidopsis Book·Dorothea Tholl, Sungbeom Lee
Apr 23, 2020·Biotechnology for Biofuels·Chien-Yuan Lin, Aymerick Eudes
May 3, 2018·FEMS Microbiology Letters·Valerie C A WardGregory Stephanopoulos
Sep 29, 2019·The Journal of Biological Chemistry·Hiroshi A Maeda
Dec 26, 2018·Proceedings of the National Academy of Sciences of the United States of America·Alkiviadis Orfefs ChatzivasileiouGregory Stephanopoulos
Nov 30, 2006·Biochemistry·Calvin N SteussyAutumn L Sutherlin
Sep 4, 2009·Biomedical and Environmental Sciences : BES·Ya-Li BenDe-Li Liu
Dec 27, 2011·Progress in Lipid Research·Andréa HemmerlinThomas J Bach

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