Characterization of Escherichia coli EutD: a phosphotransacetylase of the ethanolamine operon.
The Escherichia coli genes pta and eutD encode proteins containing the phosphate-acetyltransferase domain. EutD is composed only by this domain and belongs to the ethanolamine operon. This enzyme has not been characterized yet, and its relationship to the multimodular E. coli phosphotransacetylase (Pta) remains unclear. In the present work, a detailed characterization of EutD from E. coli (EcEutD) was performed. The enzyme is a more efficient phosphotransacetylase than E. coli Pta (EcPta) in catalyzing its reaction in either direction and assembles as a dimer, being differentially modulated by EcPta effectors. When comparing EutD and Pta, both from E. coli, certain divergent regions of the primary structure responsible for their unique properties can be found. The growth on acetate of the E. coli pta acs double-mutant strain, was complemented by either introducing EcEutD or by inducing the eut operon with ethanolamine. In this case, the expression of a phosphotransacetylase different from Pta was confirmed by activity assays. Overall, the results indicate that EcEutD and Pta, although able to catalyse the same reaction, display differential efficiency and regulation, and also differ in the induction of their expression. However, under certain growth conditions, they can fulfil equal roles in E. coli metabolism.
Centro de Estudios Fotosintéticos y Bioquímicos, Universidad Nacional de Rosario, Suipacha, 531 Rosario, Argentina.
Federico P
FP
Bologna
Valeria A
VA
Campos-Bermudez
Damián D
DD
Saavedra
Carlos S
CS
Andreo
María F
MF
Drincovich
D000085:Acetates / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D019856:Ethanolamine / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D055786:Gene Knockout Techniques; D005816:Genetic Complementation Test; D053858:Metabolic Networks and Pathways; D008954:Models, Biological; D009876:Operon; D010709:Phosphate Acetyltransferase / Q000235:genetics / Q000378:metabolism*; D055503:Protein Multimerization
D000085:Acetates; D029968:Escherichia coli Proteins; D019856:Ethanolamine; D010709:Phosphate Acetyltransferase