Three methods for the purification of Staphylococcus aureus delta haemolysin were compared (Kantor et al., 1972; Kreger et al., 1971; Heatley, 1971). The products of these purifications from the culture supernatant of S. aureus strain RN25 were compared by electrophoresis, amino acid analysis, amino-terminal sequence analysis and thin-layer chromatography. The method of Heatley (1971) was found to be superior in terms of recovery and purity of the product. Delta haemolysin prepared by the method of Kreger et al. (1971) could not be sequenced successfully prior to treatment aimed at the removal of the N-formyl group at the amino-terminus. Delta haemolysin appears to exist in two distinct molecular forms, one with N-formylmethionine and the other with un-formylated methionine in the amino-terminal position. The former polypeptide species is purified preferentially by the method of Kreger et al. (1971). Thin-layer chromatography of the products of each method of purification revealed that they were all heterogeneous, although the major component from the product of the method of Heatley (1971) represented not less than 70% of the product.
BridgenJ.,
GraffeoA.,
KargerB.L.,
WaterfieldM.1975; The identification of PTH amino acids. In Instrumentation in Amino Acid Sequence Analysis pp. 111–145PerhamR.N.
Edited by London: Academic Press;
GlazerA.N.,
DelangeR.J.,
SigmanD.S.1975; Detection reagents for peptides and amino acids. In Chemical Modification of Proteins p. 183WorkT.S.,
WorkE.
Edited by Amsterdam: North-Holland Publishing Co.;
HeatleyN.G.1976; Unusual behaviour of a protein: reversible quantitative precipitation of staphylococcal delta haemolysin by low concentrations of some organic solvents. International Journal of Peptide and Protein Research 8:233–236
KantorH.S.,
TemplesB.,
ShawW.V.1972; Staphylococcal delta haemolysin: purification and characterisation. Archives of Biochemistry and Biophysics 151:142–146
KregerA.S.,
Kwang-Shin Kim,
ZaboretskyF.,
BernheimerA.W.1971; Purification and properties of staphylococcal delta haemolysin. Infection and Immunity 3:449–465
LeeS.H.,
SarauskasT.,
HoyE.S.,
Riaz-Ul Haque.
1976; Purity of staphylococcal delta haemolysin obtained by three different procedures. Journal of Medical Microbiology 9:371–377
MachleidtW.,
WachterE.,
ScheulenM.,
OttoJ.1973; Solid-phase Edman degradation of a protein: TV-terminal sequence of cytochrome c from Candida krusei
. FEBS Letters 37:217–220
MccartneyA.C.,
ArbuthnottJ.P.1978; Mode of action of membrane-damaging toxins produced by staphylococci. In Bacterial Toxins and Cell Membranes pp. 89–127Jel-jaszewiczJ.,
WadströmT.
Edited by London: Academic Press;
SmithG.M.,
FittonJ.E.,
ShawW.V.1979; Delta haemolysin of Staphylococcus aureus: purification, structure and biosynthesis. Society for General Microbiology Quarterly 6:147