Properties of a phosphocarrier protein (HPr) extracted from intact cells of Streptococcus sanguis.
Cells of Streptococcus sanguis strain Challis were incubated with sodium lauroylsarcosinate to extract surface proteins. A polypeptide of apparent molecular mass 16 kDa comprising about 12% of the extract was purified using anion-exchange chromatography. The polypeptide was shown to be a phosphocarrier protein (HPr) that could also be found in the soluble (cytoplasmic) fraction from cells broken by homogenization with glass beads. In vivo labelling of S. sanguis cells with 32Pi showed that the polypeptide carried a heat- and acid-stable phosphorylation and that during sucrose starvation the HPr became dephosphorylated. Antiserum raised to the S. sanguis HPr reacted on Western blots with HPrs from all oral streptococci tested, together with strains of S. pyogenes and S. salivarius, but not with HPrs from S. faecalis or S. bovis, nor with proteins from Staphylococcus aureus, Bacillus subtilis, Actinomyces viscosus and various lactobacilli. The S. sanguis HPr had a high content of alanine (17.2%) and was similar in overall amino acid composition to the HPrs from S. mutans an S. salivarius. The N-terminal residues (to 37) of the S. sanguis HPr showed strong sequence identity (82%) with the N-terminal sequence of S. faecalis HPr. It is suggested that HPr in S. sanguis is associated closely with the cytoplasmic membrane. Non-disruptive methods of removing cell-surface proteins from streptococci effect release of HPr and possibly other cytoplasmic components.
Jenkinson
H F
HF
Department of Oral Biology and Oral Pathology, University of Otago, Dunedin, New Zealand.
D000595:Amino Acid Sequence; D000941:Antigens / Q000032:analysis; D001426:Bacterial Proteins / Q000302:isolation & purification*; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D008565:Membrane Proteins / Q000302:isolation & purification; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000276:immunology / Q000302:isolation & purification*; D010766:Phosphorylation; D013298:Streptococcus sanguis / Q000201:enzymology*
D000941:Antigens; D001426:Bacterial Proteins; D008565:Membrane Proteins; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; C030182:phosphocarrier protein HPr