Role of a structurally equivalent phenylalanine residue in catalysis and thermal stability of formate dehydrogenases from different sourсes

Tishkov, V.I.; Goncharenko, K.V.; Alekseeva, A.A.; Kleymenov, S.Y.; Savin, S.S.

Авторы: Tishkov V.I., Goncharenko K.V., Alekseeva A.A., Kleymenov S.Yu, Savin S.S.
Журнал: Biochemistry (Moscow)
Том: 80
Номер: 13
Год издания: 2015
Издательство: Pleiades Publishing, Ltd
Местоположение издательства: Road Town, United Kingdom
Первая страница: 1690
Последняя страница: 1700
DOI: 10.1134/S0006297915130052
Аннотация: Comparison of amino acid sequences of NAD+-dependent formate dehydrogenases (FDH, EC 1.2.1.2) from different sources and analysis of structures of holo-forms of FDH from bacterium Pseudomonas sp.101 (PseFDH) and soya Glycine max(SoyFDH) as well as of structure of apo-form of FDH from yeast Candida boidinii (CboFDH) revealed presence on the surface of protein globule of hydrophobic Phe residue in structurally equivalent position (SEP). The residue is placed in coenzyme-binding domain and protects of bound NAD+ from solvent. Comparison of influence of amino acid changes of the SEP on catalytic properties and thermal stability of PseFDH, SoyFDH и CboFDH has been carried out. The strongest effect was observed for SoyFDH. All eight amino acid replacements resulted in increase of thermal stability and in seven cases increase of catalytic constant was achieved. Thermal stability of SoyFDH after mutations Phe290Asp and Phe290Glu increased 66- and 55-fold, respectively. KM values of the enzyme for substrate and coenzyme in different cases slightly increased or decreased. In the case of CboFDH in one mutein catalytic constant increased and thermal stability did not changed. In the case of the second mutant CboFDH results were opposite. In one mutant PseFDH amino acid change did not influence on catalytic constant, but in three other ones the parameter reduced. Two mutant PseFDHs had higher and two mutants lower thermal stability in comparison with initial enzyme. Analysis of results of mutagenesis of Phe residue in structurally equivalent position in FDHs from bacterium, yeast and plant shows that protein structure play a key role for effect of the same amino acid changes in equivalent position in protein globule of formate dehydrogenases from different sources.
Добавил в систему: Тишков Владимир Иванович

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Role of a structurally equivalent phenylalanine residue in catalysis and thermal stability of formate dehydrogenases from different sourсesстатья

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