Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Roles of the Residues Lys115 and Tyr116 in the Binding of an Allosteric Inhibitor AMP to Pea Cytosolic Fructose-1,6-bisphosphatase

  • Jang, Hye-Kyung (Plant Metabolism Research Center and Graduate School of Biotechnology, Kyung Hee University) ;
  • Cho, Man-Ho (Plant Metabolism Research Center and Graduate School of Biotechnology, Kyung Hee University) ;
  • Kwon, Yong-Kook (Plant Metabolism Research Center and Graduate School of Biotechnology, Kyung Hee University) ;
  • Bhoo, Seong-Hee (Plant Metabolism Research Center and Graduate School of Biotechnology, Kyung Hee University) ;
  • Jeon, Jong-Seong (Plant Metabolism Research Center and Graduate School of Biotechnology, Kyung Hee University) ;
  • Hahn, Tae-Ryong (Plant Metabolism Research Center and Graduate School of Biotechnology, Kyung Hee University)
  • Published : 2008.04.30
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Abstract

Cytosolic fructose-1,6-bisphosphatase (cFBPase) in plants is a key regulatory enzyme in the photosynthetic sucrose biosynthesis. Plant cFBPases, like the mammalian FBPases, are inhibited by adenosine 5'-monophosphate (AMP) and fructose-2,6-bisphosphate (Fru-2,6-$P_2$). In the mammalian FBPases, Lys112 and Tyr113 play important roles in the AMP binding. To understand roles of the corresponding residues, Lys115 and Tyr116, in pea cFBPase, the mutant cFBPases were generated by site-directed mutagenesis. The alterations of Lys115 to Gin and Tyr116 to Phe displayed small changes in $K_m$ and $K_i$ for Fru-2,6-$P_2$, indicating that the mutation causes minor effects on the enzyme catalysis and Fru-2,6-$P_2$ binding, whereas resulted in higher than 500-fold increase of $[AMP]_{0.5}$ compared with that of the wild-type enzyme. Results indicate the residues Lys115 and Tyr116 play important roles in the binding of AMP to the allosteric site of the pea cFBPase.

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References

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