S layer protein of Clostridium tetani: purification and properties.
S layer protein of Clostridium tetani strain AO 174, a nontoxigenic derivative of strain Harvard A 47, was prepared from the cell walls by 4 M urea extraction and purified by DEAE-Sepharose CL-6B chromatography followed by a combination of anion-exchange chromatography and reverse-phase chromatography using an HPLC system. The molecular weight of the S layer protein was estimated to be 140 kilodaltons (kDa) by SDS-PAGE. The amino acid composition of the 140 kDa protein was very similar to those of S layer proteins from the other bacterial species: it was rich in acidic amino acid and lacked cysteine. Also, the protein was unique in its extremely low content of proline (0.02 to 0.03 mol%). Multiple isoelectric forms ranging from pH 4.0 to 4.5 were observed in the purified preparation. Immunodiffusion analysis showed that the 140 kDa protein was a common antigen to the three strains of C. tetani tested.
Takumi
K
K
Department of Food Microbiology, School of Medicine, University of Tokushima.
Susami
Y
Y
Takeoka
A
A
Oka
T
T
Koga
T
T
D000596:Amino Acids / Q000032:analysis*; D000942:Antigens, Bacterial; D001426:Bacterial Proteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D002473:Cell Wall / Q000737:chemistry; D002848:Chromatography, DEAE-Cellulose; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003017:Clostridium tetani / Q000737:chemistry*; D003545:Cysteine / Q000032:analysis; D008970:Molecular Weight; D011392:Proline / Q000032:analysis
D000596:Amino Acids; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D011392:Proline; D003545:Cysteine