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Site-directed mutagenesis of the formate dehydrogenase active centre: Role of the His(332)-G1n(313) pair in enzyme catalysisстатья
Информация о цитировании статьи получена из
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Web of Science
Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 18 июля 2013 г.
- Авторы: Tishkov V.I., Matorin A.D., Rojkova A.M., Fedorchuk V.V., Savitsky P.A., Dementieva L.A., Lamzin V.S., Mezentzev A.V., Popov V.O.
- Журнал: FEBS Letters
- Том: 390
- Номер: 1
- Год издания: 1996
- Издательство: Elsevier BV
- Местоположение издательства: Netherlands
- Первая страница: 104
- Последняя страница: 108
- DOI: 10.1016/0014-5793(96)00641-2
- Аннотация: Gln(313) and His(332) residues in the active centre of NAD(+)-dependent formate dehydrogenase (EC 1.2.1.2, FDH) from the bacterium Pseudomonas sp. 101 are conserved in all FDHs and are equivalent to the glutamate-histidine pair in active sites of D-specific 2-hydroxyacid dehydrogenases. Two mutants of formate dehydrogenase from Pseudomonas sp, 101, Gln(313) Glu and His(332)Phe, have been obtained and characterised, The Gln(313)Glu mutation shifts the pK of the group controlling formate binding from less than 5.5 in wild-type enzyme to 7.6 thus indicating that Gln(313) is essential for the broad pH affinity profile towards substrate. His(332)Phe mutation leads to a complete loss of enzyme activity. The His(332)Phe mutant is still able to bind coenzyme but not substrate or analogues, The role of histidine in the active centre of FDH is discussed, The protonation state of His(332) is not critical for catalysis but vital for substrate binding. A partial positive charge on the histidine imidazole, required for substrate binding, is provided via tight H-bond to the Gln(313) carboxamide.
- Добавил в систему: Тишков Владимир Иванович