Structure and function of proteins involved in sugar transport by the PTS of Gram-positive bacteria

Hengstenberg, Wolfgang; Reiche, B; Eisermann, Reinhard; Fischer, Roland; Tarrach, Anne; Vos, Willem M. de; Kalbitzer, Hans-Robert; Glaser, Stephen R.

doi:10.1111/j.1574-6968.1989.tb14098.x

Cited by 16 publications

(11 citation statements)

References 17 publications

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“…secondary structure motif, a four-stranded antiparallel ppleated sheet found in the NMR structure of HPr from E. coli but not in the crystal structure, was confirmed also in the NMR structures of homologeous proteins from different Gram-positive bacteria, namely in HPr from Streptococcus faecalis (Glaser, S., 1987;Hengstenberg et al, 1989;Lorenz, M., 1990), Bacillus subtilis (Wittekind et al, , 1990(Wittekind et al, , 1992 and Staphylococcus aureus (Kalbitzer et al, 1990(Kalbitzer et al, , 1991. Recently, the X-ray structures of HPr proteins from S. subtilis (Herzberg et al, 1992) and S. faecalis (Jia et al, 1993) have been solved which are in line with the NMR structures already published.…”

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confidence: 85%

The solution structure of the histidine‐containing protein (HPr) from Staphylococcus aureus as determined by two‐dimensional ¹H‐NMR spectroscopy

Kalbitzer

Hengstenberg²

1993

European Journal of Biochemistry

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The three-dimensional solution structure of the heat-stable phosphocarrier protein HPr from Staphylococcus aureus was determined from two-dimensional NMR data by restrained molecular dynamics. It consists of a large twisted antiparallel P-pleated sheet with four strands A, B, C, and D of amino acids 2-7, 34-37, 40-42 and 60-65. Three right-handed helices A, B, C (amino acids 18-27, 47-53 and 71-85) are positioned on top of this sheet. The aromatic ring of His15 is located in a cleft formed by amino acids 12-17 and 55-58, only the nitrogen (N61) atom which can be phosphorylated by enzyme I is exposed to the water. The side chains of Thrl2 and Argl7 are located close to the histidine ring. The regulatory serine residue (Ser46) is located in a hydrophobic patch, its hydroxyl group is water-accessible but forms hydrogen bonds with the amide groups of the backbone. The general features of the three-dimensional structure are similar to those found in HPr proteins from different microorganisms such as Escherichia coli, Bacillus subtilis and Streptococcus faecalis.

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confidence: 85%

The solution structure of the histidine‐containing protein (HPr) from Staphylococcus aureus as determined by two‐dimensional ¹H‐NMR spectroscopy

Kalbitzer

Hengstenberg²

1993

European Journal of Biochemistry

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“…Although first described for Escherichia coli, this group translocation system is now recognized as the primary mechanism for sugar accumulation by many species from both gram-negative (21, 26) and gram-positive bacterial genera (15,30,42,44). In a series of sequential transfers of the high-energy phosphoryl moiety from PEP, this multicomponent system catalyzes the simultaneous translocation and phosphorylation of sugars into the cell (21,26,36).…”

Section: ؉mentioning

confidence: 99%

Purification from Fusobacterium mortiferum ATCC 25557 of a 6-phosphoryl-O-alpha-D-glucopyranosyl:6-phosphoglucohydrolase that hydrolyzes maltose 6-phosphate and related phospho-alpha-D-glucosides

et al. 1995

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6-Phosphoryl-O-␣-D-glucopyranosyl:6-phosphoglucohydrolase (6-phospho-␣-glucosidase) has been purified from Fusobacterium mortiferum ATCC 25557. p-Nitrophenyl-␣-D-glucopyranoside 6-phosphate (pNP␣Glc6P) served as the chromogenic substrate for detection and assay of enzyme activity. The O 2 -sensitive, metaldependent phospho-␣-glucosidase was stabilized during purification by inclusion of dithiothreitol and Mn 2؉ ion in chromatography buffers. Various 6-phosphoryl-O-␣-linked glucosides, including maltose 6-phosphate, pNP␣Glc6P, trehalose 6-phosphate, and sucrose 6-phosphate, were hydrolyzed by the enzyme to yield D-glucose 6-phosphate and aglycone moieties in a 1:1 molar ratio. 6-Phospho-␣-glucosidase (M r of ϳ49,000; pI of ϳ4.9) is activated by Fe 2؉, Mn 2؉, Co 2؉, and Ni 2؉, and the maximum rate of pNP␣Glc6P hydrolysis occurs at 40؇C within the pH range 7.0 to 7.5. The sequence of the first 32 amino acids of 6-phospho-␣-glucosidase exhibits 67% identity (90% similarity) to that deduced for the N terminus of a putative phospho-␤-glucosidase (designated ORF f212) encoded by glvG in Escherichia coli. Western blots involving highly specific polyclonal antibody against 6-phospho-␣-glucosidase and spectrophotometric analyses with pNP␣Glc6P revealed only low levels of the enzyme in glucose-, mannose-, or fructose-grown cells of F. mortiferum. Synthesis of 6-phospho-␣-glucosidase increased dramatically during growth of the organism on ␣-glucosides, such as maltose, ␣-methylglucoside, trehalose, turanose, and palatinose.A landmark in our understanding of carbohydrate dissimilation by microorganisms was established in 1964 by the serendipitous discovery of the phosphoenolpyruvate-dependent: sugar phosphotransferase system (PEP:PTS) by Saul Roseman and his colleagues (17, 35). Although first described for Escherichia coli, this group translocation system is now recognized as the primary mechanism for sugar accumulation by many species from both gram-negative (21, 26) and gram-positive bacterial genera (15,30,42,44). In a series of sequential transfers of the high-energy phosphoryl moiety from PEP, this multicomponent system catalyzes the simultaneous translocation and phosphorylation of sugars into the cell (21,26,36).The phosphorylated products of hexose-and polyol-specific PEP:PTSs may directly enter the energy-generating (fermentation) pathways of the organism. By contrast, further metabolism of intracellular disaccharide phosphates necessitates the initial cleavage of these compounds to yield the appropriate free and phosphorylated hexose moieties. Sugar-specific (and frequently inducible) hydrolases catalyze the cleavage of the O-glycosidic linkage of the various disaccharide phosphates. Several such enzymes have been described, including those responsible for the hydrolysis of 6-phospho-␤-galactosides (6, 14, 16), 6-phospho-␤-glucosides (38, 39, 48), sucrose 6-phosphate (sucrose 6P) (7,18,46), cellobiose 6P (22), and trehalose 6P (31).Reports of maltose-PEP:PTS activity have engendered debate (5, 12, 20, 28, 41, 49),...

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“…A histidine residue (His-76) is located in a region which is highly conserved among different organisms. This residue has been proposed as the site of phosphorylation for the lactose-specific enzyme III in L. casei (1,20).…”

Section: Resultsmentioning

confidence: 99%

Cloning and sequencing of a cellobiose phosphotransferase system operon from Bacillus stearothermophilus XL-65-6 and functional expression in Escherichia coli

Lai

Ingram

1993

J Bacteriol

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Cellulolytic strains of Bacillus stearothermophilus were isolated from nature and screened for the presence of activities associated with the degradation of plant cell walls. One isolate (strain XL-65-6) which exhibited strong activities with 4-methylumbelliferyl-4-D-glucopyranoside (MUG) and 4-methylumbelliferyl-p-D-cellobiopyranoside (MUC) was used to construct a gene library in Escherichia coli. Clones degrading these model substrates were found to encode the cellobiose-specific genes of the phosphoenolpyruvate-dependent phosphotransferase system (PTS). Both MUG and MUC activities were present together, and both activities were lost concurrently during subcloning experiments. A functional E. coli ptsI gene was required for MUC and MUG activities (presumably a ptsH gene also). The DNA fragment from B. stearothermophilus contained four open reading frames which appear to form a cel operon. Intergenic stop codons for celA, celB, and celC overlapped the ribosomal binding sites of the respective downstream genes. Frameshift mutations or deletions in celA, celB, and celD were individually shown to result in a loss of MUC and MUG activities. On the basis of amino acid sequence homology and hydropathy plots of translated sequences, celA and celB were identified as encoding PTS enzyme II and celD was identified as encoding PTS enzyme III. These translated sequences were remarkably similar to their respective E. coli homologs for cellobiose transport. No reported sequences exhibited a high level of homology with the celC gene product. The predicted carboxy-terminal region for celC was similar to the corresponding region of E. coli celF, a phospho-,-glucosidase. An incomplete regulatory gene (celR) and proposed promoter sequence were located 5' to the proposed cel operon. A stem-loop resembling a rho-independent terminator was present immediately downstream from celD. These results indicate that B. stearothermophilus XL-65-6 contains a cellobiose-specific PTS for cellobiose uptake. Similar systems may be present in other gram-positive bacteria.

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Structure and function of proteins involved in sugar transport by the PTS of Gram-positive bacteria

Cited by 16 publications

References 17 publications

The solution structure of the histidine‐containing protein (HPr) from Staphylococcus aureus as determined by two‐dimensional ¹H‐NMR spectroscopy

The solution structure of the histidine‐containing protein (HPr) from Staphylococcus aureus as determined by two‐dimensional ¹H‐NMR spectroscopy

Purification from Fusobacterium mortiferum ATCC 25557 of a 6-phosphoryl-O-alpha-D-glucopyranosyl:6-phosphoglucohydrolase that hydrolyzes maltose 6-phosphate and related phospho-alpha-D-glucosides

Cloning and sequencing of a cellobiose phosphotransferase system operon from Bacillus stearothermophilus XL-65-6 and functional expression in Escherichia coli

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Structure and function of proteins involved in sugar transport by the PTS of Gram-positive bacteria

Cited by 16 publications

References 17 publications

The solution structure of the histidine‐containing protein (HPr) from Staphylococcus aureus as determined by two‐dimensional 1H‐NMR spectroscopy

The solution structure of the histidine‐containing protein (HPr) from Staphylococcus aureus as determined by two‐dimensional 1H‐NMR spectroscopy

Purification from Fusobacterium mortiferum ATCC 25557 of a 6-phosphoryl-O-alpha-D-glucopyranosyl:6-phosphoglucohydrolase that hydrolyzes maltose 6-phosphate and related phospho-alpha-D-glucosides

Cloning and sequencing of a cellobiose phosphotransferase system operon from Bacillus stearothermophilus XL-65-6 and functional expression in Escherichia coli

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The solution structure of the histidine‐containing protein (HPr) from Staphylococcus aureus as determined by two‐dimensional ¹H‐NMR spectroscopy

The solution structure of the histidine‐containing protein (HPr) from Staphylococcus aureus as determined by two‐dimensional ¹H‐NMR spectroscopy