Purification and characterization of cAMP-factor from Streptococcus agalactiae by hydrophobic interaction chromatography and chromatofocusing.
CAMP-factor from Streptococcus agalactiae (group B streptococcus) was purified 60-fold from the culture supernatant to electrophoretic homogeneity in 57% yield. The purification procedure involved ammonium sulphate precipitation, ultrafiltration, hydrophobic interaction chromatography on Octyl-Sepharose and chromatofocusing on polybuffer exchanger PBE 94. The purified CAMP-factor consists of a single polypeptide chain with an apparent molecular weight of 25 kD and an isoelectric point of 8.9. The properties of the CAMP-factor demonstrated by charge-shift electrophoresis were consistent with those of an amphiphilic polypeptide.
D000645:Ammonium Sulfate; D000906:Antibodies / Q000032:analysis; D001426:Bacterial Proteins / Q000032:analysis / Q000302:isolation & purification*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002847:Chromatography, Agarose; D004591:Electrophoresis, Polyacrylamide Gel; D006460:Hemolysin Proteins; D007525:Isoelectric Focusing; D008970:Molecular Weight; D011506:Proteins / Q000032:analysis; D013292:Streptococcus agalactiae / Q000032:analysis*; D014462:Ultrafiltration
D000906:Antibodies; D001426:Bacterial Proteins; C039092:CAMP protein, Streptococcus; D006460:Hemolysin Proteins; D011506:Proteins; D000645:Ammonium Sulfate