The phosphotransferase system (PTS) of Streptomyces coelicolor - Identification and biochemical analysis of a histidine phosphocarrier protein HPr encoded by the gene ptsH

Autor(en): Parche, S
Schmid, R
Titgemeyer, F
Stichwörter: ANTIBIOTIC PRODUCTION; BACILLUS-SUBTILIS; Biochemistry & Molecular Biology; CARBON CATABOLITE REPRESSION; COMPLETE GENOME SEQUENCE; ESCHERICHIA-COLI; GLUCOSE KINASE GENE; GRAM-POSITIVE BACTERIA; HPr; MYCOPLASMA-CAPRICOLUM; NUCLEOTIDE-SEQUENCE; phosphotransferase system; protein phosphorylation; SALMONELLA-TYPHIMURIUM; Streptomyces; sugar transport
Erscheinungsdatum: 1999
Herausgeber: WILEY
Journal: EUROPEAN JOURNAL OF BIOCHEMISTRY
Volumen: 265
Ausgabe: 1
Startseite: 308
Seitenende: 317
Zusammenfassung: 
HPr, the histidine-containing phosphocarrier protein of the bacterial phosphotransferase system (PTS) controls sugar uptake and carbon utilization in low-GC Gram-positive bacteria and in Gram-negative bacteria. We have purified HPr from Streptomyces coelicolor cell extracts. The N-terminal sequence matched the product of an S. coelicolor orf, designated ptsH, sequenced as part of the S. coelicolor genome sequencing project. The ptsH gene appears to form a monocistronic operon. Determination of the evolutionary relationship revealed that S. coelicolor HPr is equally distant to all known HPr and HPr-like proteins. The presumptive phosphorylation site around histidine 15 is perfectly conserved while a second possible phosphorylation site at serine 47 is not well-conserved. HPr was overproduced in Escherichia coli in its native form and as a histidine-tagged fusion protein. Histidine-tagged HPr was purified to homogeneity. HPr was phosphorylated by its own enzyme I (EI) and heterologously phosphorylated by EI of Bacillus subtilis and Staphylococcus aureus, respectively. This phosphoenolpyruvate-dependent phosphorylation was absent in an HPr mutant in which histidine 15 was replaced by alanine. Reconstitution of the fructose-specific PTS demonstrated that HPr could efficiently phosphorylate enzyme IIFructose. HPr-P could also phosphorylate enzyme IIGlucose of B. subtilis, enzyme IILactose of S. aureus, and IIA(Mannitol) of E. coli. ATP-dependent phosphorylation was detected with HPr kinase/phosphatase of B. subtilis. These results present the first identification of a gene of the PTS complement of S. coelicolor, providing the basis to elucidate the role(s) of HPr and the PTS in this class of bacteria.
ISSN: 00142956
DOI: 10.1046/j.1432-1327.1999.00727.x

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