Inhibition of pig phosphoenolpyruvate carboxykinase isoenzymes by 3-Mercaptopicolinic acid and novel inhibitors

Hidalgo, J. ; Latorre, P. (Universidad de Zaragoza) ; Carrodeguas, J.A. (Universidad de Zaragoza) ; Velázquez-Campoy, A. (Universidad de Zaragoza) ; Sancho, J. (Universidad de Zaragoza) ; López-Buesa, P. (Universidad de Zaragoza)

Resumen: There exist two isoforms of cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C) in pig populations that differ in a single amino acid (Met139Leu). The isoenzymes have different kinetic properties, affecting more strongly the Km and Vmax of nucleotides. They are associated to different phenotypes modifying traits of considerable economic interest. In this work we use inhibitors of phosphoenolpyruvate carboxykinase activity to search for further differences between these isoenzymes. On the one hand we have used the well-known inhibitor 3-mercaptopicolinic acid. Its inhibition patterns were the same for both isoenzymes: a three-fold decrease of the Ki values for GTP in 139Met and 139Leu (273 and 873 µM, respectively). On the other hand, through screening of a chemical library we have found two novel compounds with inhibitory effects of a similar magnitude to that of 3-mercaptopicolinic acid but with less solubility and specificity. One of these novel compounds, (N''1-({5-[1-methyl-5-(trifluoromethyl)-1H-pyrazol-3-yl]-2-thienyl}methylidene)-2, 4-dichlorobenzene-1-carbohydrazide), exhibited significantly different inhibitory effects on either isoenzyme: it enhanced threefold the apparent Km value for GTP in 139Met, whereas in 139Leu, it reduced it from 99 to 69 µM. The finding of those significant differences in the binding of GTP reinforces the hypothesis that the Met139Leu substitution affects strongly the nucleotide binding site of PEPCK-C.

Idioma: Inglés

DOI: 10.1371/journal.pone.0159002

Año: 2016

Publicado en: PloS one 11, 7 (2016), 0159002[17 pp.]
ISSN: 1932-6203

Factor impacto JCR: 2.806 (2016)

Categ. JCR: MULTIDISCIPLINARY SCIENCES rank: 15 / 63 = 0.238 (2016) - Q1 - T1

Factor impacto SCIMAGO: 1.236 - Agricultural and Biological Sciences (miscellaneous) (Q1) - Medicine (miscellaneous) (Q1) - Biochemistry, Genetics and Molecular Biology (miscellaneous) (Q1)

Financiación: info:eu-repo/grantAgreement/ES/DGA/FITE-2012-2013
Financiación: info:eu-repo/grantAgreement/ES/DGA/PI078-08
Financiación: info:eu-repo/grantAgreement/ES/MINECO/AGL2008-01487ALI
Financiación: info:eu-repo/grantAgreement/ES/MINECO/BFU2013-47064-P
Financiación: info:eu-repo/grantAgreement/ES/MINECO/BIO2014-57314-REDT

Tipo y forma: Article (Published version)

Área (Departamento): Área Bioquímica y Biolog.Mole. (Dpto. Bioq.Biolog.Mol. Celular)
Área (Departamento): Área Tecnología de Alimentos (Dpto. Produc.Animal Cienc.Ali.)