Fast purification of Clostridium sordellii cytotoxin.
We have developed a rapid method for the purification of proteins, combining titration curve analysis with a two-step column chromatographic procedure. We have used this approach to purify the cytotoxin (L toxin) from Clostridium sordellii. We have also determined the amino acid composition of this cytotoxin. This toxin has a pI value of 4.20 and an Mr of 260,000, reduction of which results in a band of Mr 43,000 on sodium dodecyl sulphate polyacrylamide gel electrophoresis. Since both the proteins of Mr 260,000 and 43,000 are recognized by the polyclonal anti-C. sordellii L toxin, which neutralizes the L toxin cytotoxicity, we propose a hexameric structure for the protein of Mr 260,000, each subunit being Mr 43,000.
Baldacini
O
O
Institut de Bactériologie, Faculté de Médecine, Strasbourg, France.
Green
G A
GA
Girardot
R
R
Rihn
B
B
Monteil
H
H
D000596:Amino Acids / Q000032:analysis; D000942:Antigens, Bacterial / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000302:isolation & purification; D001427:Bacterial Toxins / Q000276:immunology / Q000302:isolation & purification*; D002852:Chromatography, Ion Exchange; D003013:Clostridium / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007525:Isoelectric Focusing; D011487:Protein Conformation; D013194:Staining and Labeling; D014462:Ultrafiltration
D000596:Amino Acids; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D001427:Bacterial Toxins