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1.
Biol. Res ; 43(2): 191-195, 2010. ilus
Artículo en Inglés | LILACS | ID: lil-567534

RESUMEN

The functional signifcance of tyrosine 207 of Saccharomyces cerevisiae phosphoenolpyruvate carboxykinase was explored by examining the kinetic properties of the Tyr207Leu mutant. The variant enzyme retained the structural characteristics of the wild-type protein as indicated by circular dichroism, intrinsic fuorescence spectroscopy, and gel-exclusion chromatography. Kinetic analyses of the mutated variant showed a 15-fold increase in Km CO2, a 32fold decrease in Vmax, and a 6-fold decrease in Km for phosphoenolpyruvate. These results suggest that the hydroxyl group of Tyr 207 may polarize CO2 and oxaloacetate, thus facilitating the carboxylation/decarboxylation steps.


Asunto(s)
Mutación/genética , Fosfoenolpiruvato Carboxilasa/genética , Saccharomyces cerevisiae/enzimología , Tirosina/genética , Catálisis , Cromatografía en Gel , Dicroismo Circular , Regulación Enzimológica de la Expresión Génica , Regulación Fúngica de la Expresión Génica , Fosfoenolpiruvato Carboxilasa/química , Espectrometría de Fluorescencia , Tirosina/química
2.
Indian J Exp Biol ; 1991 May; 29(5): 407-10
Artículo en Inglés | IMSEAR | ID: sea-57833

RESUMEN

Transcription of phosphoenolpyruvate carboxykinase (PEPCK) gene is induced in response to cyclic AMP (cAMP) or cAMP elevating hormones. The role of transcription factors (DNA binding proteins) in the induction process has been studied. Two nuclear proteins, apparent mol. wt of 53 and 30 kDa, have been shown to bind to the 5'-flanking DNA of PEPCK gene which contains hormonal responsive elements as well as TATA box. DNA binding activity of 53 kDa protein increases by 3.5 fold in cells treated with 8-(4-chlorophenylthio)-cAMP (8-CPT-cAMP). The increased binding activity may be due to the phosphorylation of this protein by an activated cAMP-dependent protein kinase (cA kinase) in treated cells. Based on this observation, a hypothesis that 53 kDa may be specific transcription factor for PEPCK and therefore, play a major role in the regulation of this gene is proposed.


Asunto(s)
Línea Celular , Proteínas de Unión al ADN/metabolismo , Fosfoenolpiruvato Carboxilasa/genética
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