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J Bacteriol. 1971 April; 106(1): 138-142
Copyright © 1971 American Society for Microbiology. All Rights Reserved.

Ultracentrifugal Analysis of Staphylococcal Alpha Toxin

L. Forlani, A. W. Bernheimer1 and E. Chiancone

a Institute of Biochemistry and C. N. R. Center for Molecular Biology, University of Rome, Rome, Italy

ABSTRACT

Ultracentrifugal examination of staphylococcal alpha toxin at different stages of purification showed the presence of a major component having a sedimentation coefficient of 2.8S, present to the extent of more than 90% of the sample, and identifiable with active toxin. Several minor components having S20,w values of 11.5S, 8.5S, and 2.0S were detected. The 11.5S component presumably is identical with a toxin aggregate studied earlier and designated 12S; the 8.5S component appears to be delta toxin. A sedimentation equilibrium study of more highly purified material gave 32,700 as the best estimate of molecular weight of alpha toxin. Lowering the pH of the partially purified alpha toxin from 10.2 to 5.3 resulted in a small increase in S20,w of the 11.5S component and in the disappearance of the 8.5S component, whereas the S20,w, molecular weight, and hemolytic activity of the toxin remained constant. Exposure of toxin to pH 3.5 irreversibly reduced the S20,w to 2.0S, the molecular weight to about 16,000, and caused irreversible inactivation. Raising the pH of acid-inactivated toxin and adding sodium dodecyl sulfate to 1% increased the S20,w to near its normal value (2.7S) but did not restore activity.

FOOTNOTES

1 Present address: Department of Microbiology, New York University School of Medicine, New York, N.Y. 10016.

J Bacteriol. 1971 April; 106(1): 138-142
Copyright © 1971 American Society for Microbiology. All Rights Reserved.





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