Genetic evidence for the role of a bacterial phosphotransferase system in sugar transport.

Simoni, Robert D.; Levinthal, Mark; Kundig, F. Dodyk; Kündig, W.; Anderson, Byron; Hartman, Philip; Roseman, Saul

doi:10.1073/pnas.58.5.1963

Cited by 130 publications

(56 citation statements)

References 22 publications

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“…Indeed, the PTS was first discovered as a sugar "kinase" (2) and only somewhat later recognized to be a translocase (3).…”

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confidence: 99%

“…The phosphoenolpyruvate:glycose phosphotransferase system (PTS), 3 is widely distributed in bacteria and has several important roles in these cells, the most general being PTS sugar uptake where these substrates are translocated across the cytoplasmic membrane concomitant with their phosphorylation. Indeed, the PTS was first discovered as a sugar "kinase" (2) and only somewhat later recognized to be a translocase (3).…”

mentioning

confidence: 99%

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The Monomer/Dimer Transition of Enzyme I of the Escherichia coli Phosphotransferase System

Patel¹,

Vyas²,

Savtchenko

et al. 2006

Journal of Biological Chemistry

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Enzyme I (EI) is the first protein in the phosphotransfer sequence of the bacterial phosphoenolpyruvate:glycose phosphotransferase system. This system catalyzes sugar phosphorylation/transport and is stringently regulated. Since EI homodimer accepts the phosphoryl group from phosphoenolpyruvate (PEP), whereas the monomer does not, EI may be a major factor in controlling sugar uptake. Previous work from this and other laboratories (e.g. Dimitrova, M. N., Szczepanowski, R. H., Ruvinov, S. B., Peterkofsky, A., and Ginsburg A. (2002) Biochem. 41, 906 -913), indicate that K a is sensitive to several parameters. We report here a systematic study of K a determined by sedimentation equilibrium, which showed that it varied by 1000-fold, responding to virtually every parameter tested, including temperature, phosphorylation, pH (6.5 versus 7.5), ionic strength, and especially the ligands Mg 2؉ and PEP. This variability may be required for a regulatory protein. The phosphoenolpyruvate:glycose phosphotransferase system (PTS), 3 is widely distributed in bacteria and has several important roles in these cells, the most general being PTS sugar uptake where these substrates are translocated across the cytoplasmic membrane concomitant with their phosphorylation. Indeed, the PTS was first discovered as a sugar "kinase" (2) and only somewhat later recognized to be a translocase (3).The system has been extensively studied and reviewed (4, 5). Although variations of the basic motif are known, the most general phosphotransfer sequence is as follows. PEP3 Enzyme I 3 HPr 3 Sugar-specific Enzymes II SugarsEach step is physiologically reversible except for the last, phosphotransfer to the sugar. The phosphotransfer potential of PEP is 14.7 kcal/ mol, about twice that of ATP and greater than any other naturally occurring phosphate derivative. Since the phosphotransfer potentials of the PTS proteins are close to that of PEP, the energetics of the system strongly favor sugar uptake (6). From these considerations alone, it is apparent that the PTS must be stringently regulated, and indeed it is. Even the earliest results on the glucose permease by Kepes (7), before the PTS was discovered (2), showed that when a noncatabolizable Glc analogue, methyl ␣-D-glucopyranoside, is taken up by intact cells, the rate of uptake declined virtually immediately. Thus, the progress curves for uptake of PTS sugars resemble hyperbolas. These results are observed not only with intact cells but also with membrane vesicles supplied with unlimited quantities of PEP (8). We originally suggested Enzyme I as a potential candidate for governing the system (6). This idea is based on the facts that EI monomer forms a homodimer (9, 10), that the dimer but not the monomer is phosphorylated by PEP in the presence of Mg 2ϩ , and that the rate of association/dissociation is surprisingly slow, much slower than sugar uptake (11)(12)(13). This difference in rates suggests that regulation of sugar transport could be affected by factors or ligands (e.g. metabolites or other prote...

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“…Indeed, the PTS was first discovered as a sugar "kinase" (2) and only somewhat later recognized to be a translocase (3).…”

mentioning

confidence: 99%

mentioning

confidence: 99%

The Monomer/Dimer Transition of Enzyme I of the Escherichia coli Phosphotransferase System

Patel¹,

Vyas²,

Savtchenko

et al. 2006

Journal of Biological Chemistry

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“…Another example of genetic correlation are mutants defective in Enzyme I or HPr of the phosphotransferase system in E . coli or Salmonella typhimurium which have been reported to carry a pleiotropic transport defect for a number of sugars [15,16].…”

Section: Discussionmentioning

confidence: 99%

Close Linkage between a Galactose Binding Protein and the β‐Methylgalactoside Permease in Escherichia coli

Boos

Sarvas

1970

European Journal of Biochemistry

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It has been found that a gene necessary for the production of the galactose binding protein is closely linked, if not identical with the gene(s) determining the β‐methylgalactoside permease. This was determined by crossing a galactose binding protein negative, β‐methylgalactoside permease negative strain of E. coli K‐12 with wild type donor strains. In addition, after mutagenation of a permease positive, binding protein positive strain of E. coli K‐12 with N‐methyl, N‐nitroso, N′‐nitroguanidine, some of the β‐methylgalactoside permease defective mutants showed concomitant changes in the galactose binding protein. Out of 28 independent permease defective mutants two had lost their ability to produce cross reacting material against antibodies specific for the galactose binding protein; four mutants showed temperature sensitive synthesis of cross reacting material; five mutants had immunochemically somewhat changed binding protein. The immunochemical assay showed no changes in the binding protein of the rest of the 17 mutants. However, the galactose binding activity of the crude extract of these mutants varied to a great extent (10–90%) from the activity‐found in the crude extract of the wild type. These results suggest a close relationship between the galactose binding protein and the β‐methylgalactoside permease.

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“…The prokaryotic phosphoenolpyruvate (PEP)-sugar phosphotransferase system (PTS) is an unusual system for taking up sugars as it couples sugar uptake to phosphorylation (Kaback, 1968(Kaback, , 1970Lengeler & Jahreis, 2009;Postma et al, 1993;Simoni et al, 1967). It serves as an enzyme-coupled transport system that modifies its substrate sugars by phosphorylation concomitant with transport (Meadow et al, 1990).…”

Section: The Bacterial Phosphotransferase System (Pts)mentioning

confidence: 99%

Lipid dependencies, biogenesis and cytoplasmic micellar forms of integral membrane sugar transport proteins of the bacterial phosphotransferase system

Aboulwafa

Saier

2013

Microbiology

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Permeases of the prokaryotic phosphoenolpyruvate-sugar phosphotransferase system (PTS) catalyse sugar transport coupled to sugar phosphorylation. The lipid composition of a membrane determines the activities of these enzyme/transporters as well as the degree of coupling of phosphorylation to transport. We have investigated mechanisms of PTS permease biogenesis and identified cytoplasmic (soluble) forms of these integral membrane proteins. We found that the catalytic activities of the soluble forms differ from those of the membrane-embedded forms. Transport via the latter is much more sensitive to lipid composition than to phosphorylation, and some of these enzymes are much more sensitive to the lipid environment than others. While the membrane-embedded PTS permeases are always dimeric, the cytoplasmic forms are micellar, either monomeric or dimeric. Scattered published evidence suggests that other integral membrane proteins also exist in cytoplasmic micellar forms. The possible functions of cytoplasmic PTS permeases in biogenesis, intracellular sugar phosphorylation and permease storage are discussed.

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Genetic evidence for the role of a bacterial phosphotransferase system in sugar transport.

Cited by 130 publications

References 22 publications

The Monomer/Dimer Transition of Enzyme I of the Escherichia coli Phosphotransferase System

The Monomer/Dimer Transition of Enzyme I of the Escherichia coli Phosphotransferase System

Close Linkage between a Galactose Binding Protein and the β‐Methylgalactoside Permease in Escherichia coli

Lipid dependencies, biogenesis and cytoplasmic micellar forms of integral membrane sugar transport proteins of the bacterial phosphotransferase system

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