Importance of aspartate residues in balancing the flexibility and fine-tuning the catalysis of human 3-phosphoglycerate kinase

A. Varga, Zoltan Palmai, Zoltán Gugolya, Éva Gráczer, F. Vonderviszt, P. Závodszky, E. Balog, M. Vas

Research output: Contribution to journalArticle

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Abstract

The exact role of the metal ion, usually Mg2+, in the catalysis of human 3-phosphoglycerate kinase, a well-studied two-domain enzyme, has not been clarified. Here we have prepared single and double alanine mutants of the potential metal-binding residues, D374 and D218. While all mutations weaken the catalytic interactions with Mg2+, they surprisingly strengthen binding of both MgADP and MgATP, and the effects are even more pronounced for ADP and ATP. Thermodynamic parameters of binding indicate an increase in the binding entropy as a reason for the strengthening. In agreement with the experimental results, computer-simulated annealing calculations for the complexes of these mutants have supported the mobility of the nucleotide phosphates and, as a consequence, formation of their new interaction(s) within the active site. A similar type of mobility is suggested to be a characteristic feature of the nucleotide site of the wild-type enzyme, too, both in its inactive open conformation and in the active closed conformation. This mobility of the nucleotide phosphates that is regulated by the aspartate side chains of D218 and D374 through the complexing Mg2+ is suggested to be essential in enzyme function.

Original languageEnglish
Pages (from-to)10197-10207
Number of pages11
JournalBiochemistry
Volume51
Issue number51
DOIs
Publication statusPublished - Dec 21 2012

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Phosphoglycerate Kinase
Catalysis
Aspartic Acid
Nucleotides
Tuning
Adenosine Diphosphate
Conformations
Enzymes
Adenosine Triphosphate
Metals
Phosphates
Entropy
Simulated annealing
Thermodynamics
Alanine
Metal ions
Catalytic Domain
Ions
Mutation
3-phosphoglycerate

ASJC Scopus subject areas

  • Biochemistry

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Importance of aspartate residues in balancing the flexibility and fine-tuning the catalysis of human 3-phosphoglycerate kinase. / Varga, A.; Palmai, Zoltan; Gugolya, Zoltán; Gráczer, Éva; Vonderviszt, F.; Závodszky, P.; Balog, E.; Vas, M.

In: Biochemistry, Vol. 51, No. 51, 21.12.2012, p. 10197-10207.

Research output: Contribution to journalArticle

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AU - Palmai, Zoltan

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AU - Gráczer, Éva

AU - Vonderviszt, F.

AU - Závodszky, P.

AU - Balog, E.

AU - Vas, M.

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