Direct appraisal of the potato tuber ADP-glucose pyrophosphorylase large subunit in enzyme function by study of a novel mutant form

The Journal of Biological Chemistry
Seon-Kap HwangThomas W Okita

Abstract

The higher plant ADP-glucose pyrophosphorylase is a heterotetramer consisting of two subunit types, which have evolved at different rates from a common ancestral gene. The potato tuber small subunit (SS) displays both catalytic and regulatory properties, whereas the exact role of the large subunit (LS), which contains substrate and effector binding sites, remains unresolved. We identified a mutation, S302N, which increased the solubility of the recombinant potato tuber LS and, in turn, enabling it to form a homotetrameric structure. The LS302N homotetramer possesses very little enzyme activity at a level 100-fold less than that seen for the unactivated SS homotetramer. Unlike the SS enzyme, however, the LS302N homotetramer enzyme is neither activated by the effector 3-phosphoglycerate nor inhibited by P(i). When combined with the catalytically silenced SS, S D143N, however, the LS302N-containing enzyme shows significantly enhanced catalytic activity and restored 3-PGA activation. This unmasking of catalytic and regulatory potential of the LS is conspicuously evident when the activities of the resurrected L(K41R.T51K.S302N) homotetramer are compared with its heterotetrameric form assembled with S D143N. Overall, these results in...Continue Reading

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Citations

Sep 10, 2009·The Journal of Biological Chemistry·Misty L KuhnMiguel A Ballicora
Jul 11, 2008·Plant Physiology·Tiziana VentrigliaJosé M Romero
Oct 31, 2009·PLoS Computational Biology·Ibrahim BarisIbrahim Halil Kavakli
Sep 11, 2013·Bioscience, Biotechnology, and Biochemistry·Shinji WakutaHirokazu Matsui
Jun 4, 2014·Plant & Cell Physiology·A Bengisu SeferogluI Halil Kavakli
Jul 19, 2011·Plant Science : an International Journal of Experimental Plant Biology·Kelly GibsonThomas W Okita
Mar 19, 2013·Plant Science : an International Journal of Experimental Plant Biology·Ayse Bengisu SeferogluIbrahim Halil Kavakli
Jun 15, 2016·Plant Science : an International Journal of Experimental Plant Biology·Xiao-Jie TangJian-Min Wan
Oct 9, 2016·Plant Science : an International Journal of Experimental Plant Biology·Ayse Bengisu SeferogluIbrahim Halil Kavakli
May 19, 2018·Protoplasma·Asena GorenIan J Tetlow
Feb 26, 2019·Frontiers in Plant Science·Seon-Kap HwangThomas W Okita

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