PMID: 9531508Jun 11, 1998Paper

Cystathionine gamma-synthase from Arabidopsis thaliana: purification and biochemical characterization of the recombinant enzyme overexpressed in Escherichia coli

The Biochemical Journal
S RavanelR Douce

Abstract

Cystathionine gamma-synthase catalyses the first reaction specific for methionine biosynthesis in plants, the gamma-replacement of the phosphoryl substituent of O-phosphohomoserine by cysteine. A cDNA encoding cystathionine gamma-synthase from Arabidopsis thaliana has been cloned and used to overexpress the enzyme in Escherichia coli. The native recombinant enzyme is a homotetramer composed of 53 kDa subunits, each being tightly associated with one molecule of pyridoxal 5'-phosphate that binds at lysine-379 of the protein precursor. The replacement reaction follows a Ping Pong mechanism with a Vmax of 33.6 units/mg and Km values of 2.5 mM and 460 microM for O-phosphohomoserine and cysteine respectively. The protective effect of O-phosphohomoserine against enzyme inactivation by propargylglycine indicated that the Kd for the substrate is approx. 1/2500 of its Km value. Thus most of these biochemical properties are similar to those previously reported for plant and bacterial cystathionine gamma-synthases. However, the plant enzyme differs markedly from its enterobacterial counterparts because it catalyses a very faint gamma-elimination of O-phosphohomoserine in the absence of cysteine, this process being about 1/2700 as fast as t...Continue Reading

Citations

May 5, 2001·Annual Review of Plant Physiology and Plant Molecular Biology·Arthur Grossman, Hideki Takahashi
Sep 17, 2005·Biological Chemistry·Gad GaliliHolger Hesse
Nov 25, 2005·Photosynthesis Research·Markus Wirtz, Michel Droux
Sep 21, 2000·The Plant Journal : for Cell and Molecular Biology·S MaimannH Hesse
Mar 27, 2013·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Dominique J K MorneauSusan M Aitken
Jun 24, 1998·Proceedings of the National Academy of Sciences of the United States of America·S RavanelR Douce
Oct 2, 2002·Physiologia Plantarum·Karine GallardoDominique Job
May 3, 2003·FEMS Microbiology Letters·Yasuo YoshidaYoshio Nakano
Jan 1, 2009·The Arabidopsis Book·Georg Jander, Vijay Joshi
Jun 9, 2020·Pest Management Science·Cody J HallTatiana P Soares da Costa
Dec 12, 2020·Computational and Structural Biotechnology Journal·Kwanjeera WanichthanarakMaysaya Thitisaksakul
Aug 28, 2020·Plant Physiology and Biochemistry : PPB·Salvador González-GordoFrancisco J Corpas
Dec 8, 2004·Archives of Biochemistry and Biophysics·Susan M Aitken, Jack F Kirsch
Aug 7, 2021·Frontiers in Plant Science·Stefania AstolfiStefano Cesco
May 12, 2009·International Journal of Biological Macromolecules·Baisakhee SahaAmit Kumar Das

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