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Purification and Immunochemical Properties of a Protein Antigen from Serotype g Streptococcus mutans
Author(s) -
Okahashi Nobuo,
Koga Toshihiko,
Hamada Shigeyuki
Publication year - 1986
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1986.tb00919.x
Subject(s) - isoelectric point , chromatography , streptococcus mutans , size exclusion chromatography , antigen , antigenicity , immunodiffusion , radial immunodiffusion , ammonium sulfate precipitation , lipoteichoic acid , biology , biochemistry , serotype , ion chromatography , microbiology and biotechnology , chemistry , bacteria , antibody , genetics , staphylococcus aureus , immunology , enzyme
A proteinaceous antigen (PA g ) was purified from the culture supernatant of Streptococcus mutans 6715 (serotype g ) by ultrafiltration, ammonium sulfate precipitation, DEAE‐Sephacel ion‐exchange chromatography, Phenyl‐Sepharose CL‐4B hydrophobic chromatography, and subsequent Sephacryl S‐300 gel filtration. A yield of 0.1 mg of PA g was obtained from a liter of culture supernatant. The isoelectric point and molecular weight of PA g were pH 4.6 and 210,000, respectively. It contained 35% sugar, which was identified as glucose by gas‐liquid chromatography. Amino acid analysis revealed that PA g contains 28% acidic and 11% basic amino acid residues. PA g retained its antigenicity after heating at 80 C for 10 min in deionized water, or after treatment with 0.1 M HCl or 0.1 M NaOH at 37 C for 1 hr. Immunodiffusion and Immunoelectrophoresis analyses revealed that PA g is serologically distinct from other cell‐surface antigens such as serotype‐specific polysaccharide and lipoteichoic acid. A cross‐reaction between PA g and a protein antigen similarly prepared from serotype c S. mutans was observed in immunodiffusion tests.