Purification and Some Chemical and Physical Properties of Staphylococcal Enterotoxin A
Abstract
Enterotoxin A from a selected high toxin producing strain (15-25 micrograms/ml in deep culture) of Staphylococcus aureus, designated 13N-2909, has been obtained in highly purified form as indicated by sedimentation velocity, sedimentation equilibrium, disc electrophoresis, and several immunological procedures. Purification was accomplished by the following STEPS: (1) removal of the toxin from culture (diluted five times with water, and adjusted to pH 5.6) with CG-50 carboxylic acid resin; (2) chromatography on carboxymethylcellulose; (3) chromatography on hydroxlapatite; and (4) filtration on Sephadex G-75. Two major and two minor components were found by isoelectric focusing. The enterotoxin has a molecular weight of 27,800 as determined by sedimentation equilibrium. A value of 27,500 was obtained from polyacrylamide gel electrophoresis in the presence of denaturant. The molecule is a single polypeptide chain with one disulfide bridge and no free sulfhydryl groups. Serine was identified as the C-terminal residue but no free N terminal was found. A complete amino acid analysis is reported.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 02, 1971
- Accession Number
- AD0737903
Entities
People
- Edward J. Schantz
- Henry M. Jacoby
- Joseph M. Lynch
- Margy J. Woodburn
- William G. Roessler