1987
DOI: 10.1128/jb.169.6.2810-2818.1987
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Characterization of mutant histidine-containing proteins of the phosphoenolpyruvate:sugar phosphotransferase system of Escherichia coli and Salmonella typhimurium

Abstract: Histidine-containing phosphocarrier protein (HPr) is common to all of the phosphoenolpyruvate:sugar phosphotransferase systems (PTS) in Escherichia coli and SalmoneUa typhimurium, except the fructose-specific PTS. Strains which lack HPr activity (ptsH) have been characterized in the past, and it has proved difficult to delineate between tight and leaky mutants. In this study four different parameters of ptsH strains were measured: in vitro sugar phosphorylation activity of the mutant HPr; detection of 32P-labe… Show more

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Cited by 23 publications
(24 citation statements)
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“…1), were constructed and inserted into the low-expression vector, plasmid pT7-4 (43). An HPr-negative E. coli strain, LBG1650 (2,45), was then transformed with these plasmids, and the fermentation responses were recorded (Table 1). Replacement of Asp-69 with Tyr (D69Y) resulted in a marked decrease in growth rate and fermentation response on plates containing glucose, mannitol, N-acetylglucosamine, or any one of a number of other sugar substrates of the PTS.…”
Section: Resultsmentioning
confidence: 99%
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“…1), were constructed and inserted into the low-expression vector, plasmid pT7-4 (43). An HPr-negative E. coli strain, LBG1650 (2,45), was then transformed with these plasmids, and the fermentation responses were recorded (Table 1). Replacement of Asp-69 with Tyr (D69Y) resulted in a marked decrease in growth rate and fermentation response on plates containing glucose, mannitol, N-acetylglucosamine, or any one of a number of other sugar substrates of the PTS.…”
Section: Resultsmentioning
confidence: 99%
“…Two examples of mutant HPrs with hydrolysis rates that are 50% of the wild-type E. coli HPr rate have been reported. The first example is a mutant E. coli HPr (Term-85) that lacks E-85 and possesses L-84 at the C terminus, while the second example is the Enterococcus faecalis HPr, which has a C-terminal glutamine (11)(12)(13)(44)(45)(46)(47). In the case of Term-85, determination of its structure by NMR revealed an adjustment such that E-83 contributes its carboxyl group to the active site (1,11), presumably in a less effective arrangement.…”
Section: Discussionmentioning
confidence: 99%
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“…The proteins and enzymes were expressed in strains that contained suitable mutations. HPr mutants were overproduced in a ptsH strain in which a terminator codon occurs at residue 71 4 and in which no HPr activity or protein can be detected (38). HPr preparations could not be phosphorylated without the addition of enzyme I.…”
Section: Replacement Of Phosphorylated His With Acidic Amino Acidsmentioning
confidence: 99%