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Biochemical, Kinetic, and Computational Structural Characterization of Shikimate Kinase from Methicillin-Resistant Staphylococcus aureus

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Abstract

One of the most widespread pathogens worldwide is methicillin-resistant Staphylococcus aureus, a bacterium that provokes severe life-threatening illnesses both in hospitals and in the community. The principal challenge lies in the resistance of MRSA to current treatments, which encourages the study of different molecular targets that could be used to develop new drugs against this infectious agent. With this goal, a detailed characterization of shikimate kinase from this microorganism (SaSK) is described. The results showed that SaSK has a Km of 0.153 and 224 µM for shikimate and ATP, respectively, and a global reaction rate of 13.4 µmol/min/mg; it is suggested that SaSK utilizes the Bi–Bi Ping Pong reaction mechanism. Furthermore, the physicochemical data indicated that SaSK is an unstable, hydrophilic, and acidic protein. Finally, structural information showed that SaSK presented folding that is typical of its homologous counterparts and contains the typical domains of this family of proteins. Amino acids that have been shown to be important for SaSK protein function are conserved. Therefore, this study provides fundamental information that may aid in the design of inhibitors that could be used to develop new antibacterial agents.

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Acknowledgements

C.A.D. and A.T.V. acknowledge Consejo Nacional de Ciencia y Tecnología (CONACyT) for Grants Nos. 258694 and 257848. CONACyT is also acknowledged for the fellowship granted to A.F.C. (No. 326106).

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Authors and Affiliations

  1. Facultad de Medicina y Nutrición, Universidad Juárez del Estado de Durango, Av. Universidad y Fanny Anitua S/N, C.P. 34000, Durango, Dgo, Mexico

    Alejandro Favela-Candia, Alfredo Téllez-Valencia, Mara Campos-Almazán & Claudia Avitia-Domínguez

  2. Facultad de Ciencias Químicas, Universidad Juárez del Estado de Durango, Av. Artículo 123 S/N Fracc. Filadelfia, Gómez Palacio, C.P. 35010, Durango, Mexico

    Erick Sierra-Campos & Mónica Valdez-Solana

  3. Laboratorio de Bioquímica Genética, Secretaría de Salud, Instituto Nacional de Pediatría, C.P. 04534, Ciudad de México, Mexico

    Jesús Oria-Hernández & Adriana Castillo-Villanueva

  4. Departamento de Ciencias Naturales, Universidad Autónoma Metropolitana, Unidad Cuajimalpa, Delegación Cuajimalpa de Morelos, Av. Vasco de Quiroga 4871, Colonia Santa Fe Cuajimalpa, C.P. 05300, Ciudad de México, Mexico

    Hugo Nájera

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  1. Alejandro Favela-Candia
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Correspondence to Claudia Avitia-Domínguez.

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Favela-Candia, A., Téllez-Valencia, A., Campos-Almazán, M. et al. Biochemical, Kinetic, and Computational Structural Characterization of Shikimate Kinase from Methicillin-Resistant Staphylococcus aureus. Mol Biotechnol 61, 274–285 (2019). https://doi.org/10.1007/s12033-019-00159-5

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