Insights into the interaction of human liver arginase with tightly and weakly bound manganese ions by chemical modification and site-directed mutagenesis studies

María S. Orellana, Vasthi López, Elena Uribe, Marcia Fuentes, Mónica Salas, Nelson Carvajal

Resultado de la investigación: Contribución a una revistaArtículorevisión exhaustiva

10 Citas (Scopus)

Resumen

Diethyl pyrocarbonate (DEPC) caused a loss in the ability of inactive subunits of wild-type and H141F mutant human liver arginase (EC 3.5.3.1) to be reactivated by Mn2+. The effect was reversed by hydroxylamine and involved a residue with a pKa of 6.5 ± 0.1. Half activation with Mn2+ was sufficient for total resistance of H141F and full activation was not impeded by a previous incubation of the half-active species with DEPC. The H101N and H126N mutants expressed 60 and 82% of the wild-type activity, respectively, without changes in Km for arginine or Ki for lysine inhibition. After dialysis against EDTA, H126N was inactive in the absence of added Mn2+ and contained < 0.1 Mn2+/subunit, whereas H101N was half active and contained 1.2 ± 0.1 Mn2+/subunit. Results support the concept that a weakly bound metal ion is needed only for conversion of active species to a more active active state.

Idioma originalInglés
Páginas (desde-hasta)155-159
Número de páginas5
PublicaciónArchives of Biochemistry and Biophysics
Volumen403
N.º2
DOI
EstadoPublicada - 15 jul 2002
Publicado de forma externa

Áreas temáticas de ASJC Scopus

  • Biofísica
  • Bioquímica
  • Biología molecular

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