• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of plntphysLink to Publisher's site
Plant Physiol. Nov 1996; 112(3): 1315–1320.
PMCID: PMC158059

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

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 acid 413 to alanine. Substitution of a lysine residue at this site almost completely abolished activation by 3-PGA. Aspartic acid 413 is adjacent to a lysine residue that was previously identified by chemical modification studies to be important in the binding of 3-PGA (K. Ball, J. Preiss [1994] J Biol Chem 269: 24706-24711). The kinetic properties of M27 corroborate the importance of this region in the allosteric regulation of a higher-plant AGP.

Full Text

The Full Text of this article is available as a PDF (619K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Ball K, Preiss J. Allosteric sites of the large subunit of the spinach leaf ADPglucose pyrophosphorylase. J Biol Chem. 1994 Oct 7;269(40):24706–24711. [PubMed]
  • Ballicora MA, Laughlin MJ, Fu Y, Okita TW, Barry GF, Preiss J. Adenosine 5'-diphosphate-glucose pyrophosphorylase from potato tuber. Significance of the N terminus of the small subunit for catalytic properties and heat stability. Plant Physiol. 1995 Sep;109(1):245–251. [PMC free article] [PubMed]
  • Dickinson DB, Preiss J. Presence of ADP-Glucose Pyrophosphorylase in Shrunken-2 and Brittle-2 Mutants of Maize Endosperm. Plant Physiol. 1969 Jul;44(7):1058–1062. [PMC free article] [PubMed]
  • Govons S, Vinopal R, Ingraham J, Preiss J. Isolation of mutants of Escherichia coli B altered in their ability to synthesize glycogen. J Bacteriol. 1969 Feb;97(2):970–972. [PMC free article] [PubMed]
  • Hannah LC, Nelson OE., Jr Characterization of ADP-glucose pyrophosphorylase from shrunken-2 and brittle-2 mutants of maize. Biochem Genet. 1976 Aug;14(7-8):547–560. [PubMed]
  • Iglesias AA, Barry GF, Meyer C, Bloksberg L, Nakata PA, Greene T, Laughlin MJ, Okita TW, Kishore GM, Preiss J. Expression of the potato tuber ADP-glucose pyrophosphorylase in Escherichia coli. J Biol Chem. 1993 Jan 15;268(2):1081–1086. [PubMed]
  • Lin TP, Caspar T, Somerville C, Preiss J. Isolation and Characterization of a Starchless Mutant of Arabidopsis thaliana (L.) Heynh Lacking ADPglucose Pyrophosphorylase Activity. Plant Physiol. 1988 Apr;86(4):1131–1135. [PMC free article] [PubMed]
  • Lin TP, Caspar T, Somerville CR, Preiss J. A Starch Deficient Mutant of Arabidopsis thaliana with Low ADPglucose Pyrophosphorylase Activity Lacks One of the Two Subunits of the Enzyme. Plant Physiol. 1988 Dec;88(4):1175–1181. [PMC free article] [PubMed]
  • Morell M, Bloom M, Preiss J. Affinity labeling of the allosteric activator site(s) of spinach leaf ADP-glucose pyrophosphorylase. J Biol Chem. 1988 Jan 15;263(2):633–637. [PubMed]
  • Müller-Röber B, Sonnewald U, Willmitzer L. Inhibition of the ADP-glucose pyrophosphorylase in transgenic potatoes leads to sugar-storing tubers and influences tuber formation and expression of tuber storage protein genes. EMBO J. 1992 Apr;11(4):1229–1238. [PMC free article] [PubMed]
  • Nakata PA, Greene TW, Anderson JM, Smith-White BJ, Okita TW, Preiss J. Comparison of the primary sequences of two potato tuber ADP-glucose pyrophosphorylase subunits. Plant Mol Biol. 1991 Nov;17(5):1089–1093. [PubMed]
  • Okita TW. Is there an alternative pathway for starch synthesis? Plant Physiol. 1992 Oct;100(2):560–564. [PMC free article] [PubMed]
  • Okita TW, Nakata PA, Anderson JM, Sowokinos J, Morell M, Preiss J. The Subunit Structure of Potato Tuber ADPglucose Pyrophosphorylase. Plant Physiol. 1990 Jun;93(2):785–790. [PMC free article] [PubMed]
  • Smith-White BJ, Preiss J. Comparison of proteins of ADP-glucose pyrophosphorylase from diverse sources. J Mol Evol. 1992 May;34(5):449–464. [PubMed]
  • Tsai CY, Nelson OE. Starch-deficient maize mutant lacking adenosine dephosphate glucose pyrophosphorylase activity. Science. 1966 Jan 21;151(3708):341–343. [PubMed]

Articles from Plant Physiology are provided here courtesy of American Society of Plant Biologists

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Compound
    Compound
    PubChem Compound links
  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...