Aspartic acid 413 is important for the normal allosteric functioning of ADP-glucose pyrophosphorylase

Plant Physiology
Thomas W GreeneT W Okita

Abstract

As part of a structure-function analysis of the higher-plant ADP-glucose pyrophosphorylase (AGP), we used a random mutagenesis approach in combination with a novel bacterial complementation system to isolate over 100 mutants that were defective in glycogen production (T.W. Greene, S.E. Chantler, M.L. Khan, G.F. Barry, J. Preiss, T.W. Okita [1996] Proc Natl Acad Sci USA 93: 1509-1513). One mutant of the large subunit M27 was identified by its capacity to only partially complement a mutation in the structural gene for the bacterial AGP (glg C), as determined by its light-staining phenotype when cells were exposed to l3 vapors. Enzyme-linked immunosorbent assay and enzymatic pyrophosphorylysis assays of M27 cell extracts showed that the level of expression and AGP activity was comparable to those of cells that expressed the wild-type recombinant enzyme. Kinetic analysis indicated that the M27 AGP displays normal Michaelis constant values for the substrates glucose-1-phosphate and ATP but requires 6- to 10-fold greater levels of 3-phosphoglycerate (3-PGA) than the wild-type recombinant enzyme for maximum activation. DNA sequence analysis showed that M27 contains a single point mutation that resulted in the replacement of aspartic a...Continue Reading

Citations

May 4, 2004·Plant Physiology·Joanna M CrossL Curtis Hannah
Nov 6, 2009·Plant Physiology·Susan K BoehleinJon D Stewart
Jun 10, 2003·Microbiology and Molecular Biology Reviews : MMBR·Miguel A BallicoraJack Preiss
Aug 26, 1998·Proceedings of the National Academy of Sciences of the United States of America·T W GreeneT W Okita
Oct 28, 1998·Proceedings of the National Academy of Sciences of the United States of America·T W Greene, L C Hannah
Jun 4, 2014·Plant & Cell Physiology·A Bengisu SeferogluI Halil Kavakli
Feb 14, 1998·Phytochemistry·M J LaughlinT W Okita
Feb 5, 2005·The EMBO Journal·Xiangshu JinJames H Geiger
Oct 13, 2004·Protein Expression and Purification·Seon-Kap HwangThomas W Okita
Nov 15, 2018·Frontiers in Plant Science·Carlos M FigueroaMiguel A Ballicora

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