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Kinetic characterization of Vibrio cholerae ApbE: Substrate specificity and regulatory mechanisms.

ABSTRACT: ApbE is a member of a novel family of flavin transferases that incorporates flavin mononucleotide (FMN) to subunits of diverse respiratory complexes, which fulfill important homeostatic functions. In this work a detailed characterization of Vibrio cholerae ApbE physiologic activity, substrate specificity and pH dependency was carried out. The data obtained show novel characteristics of the regulation and function of this family. For instance, our experiments indicate that divalent cations are essential for ApbE function, and that the selectivity depends largely on size and the coordination sphere of the cation. Our data also show that ApbE regulation by pH, ADP and potassium is an important mechanism that enhances the adaptation, survival and colonization of V. cholerae in the small intestine. Moreover, studies of the pH-dependency of the activity show that the reaction is favored under alkaline conditions, with a pKa of 8.4. These studies, together with sequence and structure analysis allowed us to identify His257, which is absolutely conserved in the family, as a candidate for the residue whose deprotonation controls the activity. Remarkably, the mutant H257G abolished the flavin transfer activity, strongly indicating that this residue plays an important role in the catalytic mechanism of ApbE.

SUBMITTER: Fang X

PROVIDER: S-EPMC5655446 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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Kinetic characterization of Vibrio cholerae ApbE: Substrate specificity and regulatory mechanisms.

Fang Xuan X Liang Pingdong P Raba Daniel Alexander DA Rosas-Lemus Mónica M Chakravarthy Srinivas S Tuz Karina K Juárez Oscar O

PloS one 20171024 10

ApbE is a member of a novel family of flavin transferases that incorporates flavin mononucleotide (FMN) to subunits of diverse respiratory complexes, which fulfill important homeostatic functions. In this work a detailed characterization of Vibrio cholerae ApbE physiologic activity, substrate specificity and pH dependency was carried out. The data obtained show novel characteristics of the regulation and function of this family. For instance, our experiments indicate that divalent cations are es ...[more]

PMID: 29065131

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Kinetic characterization of Vibrio cholerae ApbE: Substrate specificity and regulatory mechanisms.

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Kinetic characterization of Vibrio cholerae ApbE: Substrate specificity and regulatory mechanisms.

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