Mutagenesis of the glucose-1-phosphate-binding site of potato tuber ADP-glucose pyrophosphorylase

Plant Physiology
Y FuJ Preiss

Abstract

Lysine (Lys)-195 in the homotetrameric ADP-glucose pyrophosphorylase (ADPGlc PPase) from Escherichia coli was shown previously to be involved in the binding of the substrate glucose-1-phosphate (Glc-1-P). This residue is highly conserved in the ADPGlc PPase family. Site-directed mutagenesis was used to investigate the function of this conserved Lys residue in the large and small subunits of the heterotetrameric potato (Solanum tuberosum) tuber enzyme. The apparent affinity for Glc-1-P of the wild-type enzyme decreased 135- to 550-fold by changing Lys-198 of the small subunit to arginine, alanine, or glutamic acid, suggesting that both the charge and the size of this residue influence Glc-1-P binding. These mutations had little effect on the kinetic constants for the other substrates (ATP and Mg2+ or ADP-Glc and inorganic phosphate), activator (3-phosphoglycerate), inhibitor (inorganic phosphate), or on the thermal stability. Mutagenesis of the corresponding Lys (Lys-213) in the large subunit had no effect on the apparent affinity for Glc-1-P by substitution with arginine, alanine, or glutamic acid. A double mutant, SK198RLK213R, was also obtained that had a 100-fold reduction of the apparent affinity for Glc-1-P. The data indic...Continue Reading

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Citations

Mar 10, 2001·Biochemical and Biophysical Research Communications·I H KavakliT W Okita
Oct 12, 2005·Plant Molecular Biology·Meijuan Duan, Samuel S M Sun
Apr 20, 2001·Biochimica Et Biophysica Acta·I J KimW I Chung
Dec 15, 2000·Current Opinion in Structural Biology·M F Giraud, J H Naismith
Aug 7, 2002·Proceedings of the National Academy of Sciences of the United States of America·Antonius KollerJohn R Yates
Dec 2, 2006·FEBS Letters·Seon-Kap HwangThomas W Okita
Dec 1, 2015·Frontiers in Microbiology·Ana C EbrechtMiguel A Ballicora
Feb 5, 2005·The EMBO Journal·Xiangshu JinJames H Geiger
Feb 3, 2005·The Plant Journal : for Cell and Molecular Biology·Joanna M CrossL Curtis Hannah
Oct 13, 2004·Protein Expression and Purification·Seon-Kap HwangThomas W Okita
Jul 28, 2004·Phytochemistry·Valérie MéchinCatherine Damerval
May 11, 2005·Biochemical and Biophysical Research Communications·Zhanmin FanZhangliang Chen
Jan 6, 2005·The Journal of Biological Chemistry·Miguel A BallicoraJack Preiss
Sep 17, 1998·The Journal of Biological Chemistry·Y FuJ Preiss
Jun 10, 2003·Microbiology and Molecular Biology Reviews : MMBR·Miguel A BallicoraJack Preiss
Feb 28, 2006·Plant & Cell Physiology·Alberto A IglesiasJack Preiss
Nov 3, 2006·The Journal of Biological Chemistry·Clarisa Maria BejarJack Preiss
Sep 10, 2009·The Journal of Biological Chemistry·Misty L KuhnMiguel A Ballicora
Sep 22, 2001·The Journal of Biological Chemistry·J B FrueaufJ Preiss
Jan 8, 2000·The Journal of Biological Chemistry·M A BallicoraJ Preiss
Mar 22, 2008·Biochemistry·Jill R Cupp-VickeryChristopher R Meyer
Apr 20, 1999·Biochemical and Biophysical Research Communications·M A BallicoraJ Preiss
May 1, 2014·EcoSal Plus·Jack Preiss
Aug 1, 2009·EcoSal Plus·Jack Preiss

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