Comparison of Three Methods for the Purification of the Delta Haemolysin of Free

Abstract

Three methods for the purification of delta haemolysin were compared ( Kantor ., 1972 Kreger ., 1971 Heatley, 1971 ). The products of these purifications from the culture supernatant of 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 . (1971) could not be sequenced successfully prior to treatment aimed at the removal of the -formyl group at the amino-terminus. Delta haemolysin appears to exist in two distinct molecular forms, one with -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 . (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.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-124-2-365
1981-06-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/124/2/mic-124-2-365.html?itemId=/content/journal/micro/10.1099/00221287-124-2-365&mimeType=html&fmt=ahah

References

  1. Ambler R.P. 1975; The amino acid sequence of Staphylococcus aureus penicillinase. Biochemical Journal 151:197–218
    [Google Scholar]
  2. Bernheimer A.W. 1974; Interactions between membranes and cytolytic bacterial toxins. Biochimica et biophysica acta 344:27–50
    [Google Scholar]
  3. Bridgen J., Graffeo A., Karger B.L., Waterfield M. 1975; The identification of PTH amino acids. In Instrumentation in Amino Acid Sequence Analysis pp. 111–145 Perham R.N. Edited by London: Academic Press;
    [Google Scholar]
  4. Fitton J.E., Dell A., Shaw W.V. 1980; The amino acid sequence of the delta haemolysin of Staphylococcus aureus . FEBS Letters 115:209–212
    [Google Scholar]
  5. Gladstone G, Van Heyningen W.E. 1957; Staphylococcal leucocidins. British Journal of Experimental Pathology 38:123–137
    [Google Scholar]
  6. Glazer A.N., Delange R.J., Sigman D.S. 1975; Detection reagents for peptides and amino acids. In Chemical Modification of Proteins p. 183 Work T.S., Work E. Edited by Amsterdam: North-Holland Publishing Co.;
    [Google Scholar]
  7. Heatley N.G. 1971; A new method for the preparation and some properties of staphylococcal delta-haemolysin. Journal of General Microbiology 69:269–278
    [Google Scholar]
  8. Heatley N.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
    [Google Scholar]
  9. Kantor H.S., Temples B., Shaw W.V. 1972; Staphylococcal delta haemolysin: purification and characterisation. Archives of Biochemistry and Biophysics 151:142–146
    [Google Scholar]
  10. Kreger A.S., Kwang-Shin Kim, Zaboretsky F., Bernheimer A.W. 1971; Purification and properties of staphylococcal delta haemolysin. Infection and Immunity 3:449–465
    [Google Scholar]
  11. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  12. Laursen R.A. 1971; Solid-phase Edman degradation. An automatic peptide sequencer. European Journal of Biochemistry 20:89–102
    [Google Scholar]
  13. Lee S.H., Sarauskas T., Hoy E.S., Riaz-Ul Haque. 1976; Purity of staphylococcal delta haemolysin obtained by three different procedures. Journal of Medical Microbiology 9:371–377
    [Google Scholar]
  14. Machleidt W., Wachter E., Scheulen M., Otto J. 1973; Solid-phase Edman degradation of a protein: TV-terminal sequence of cytochrome c from Candida krusei . FEBS Letters 37:217–220
    [Google Scholar]
  15. Mccartney A.C., Arbuthnott J.P. 1978; Mode of action of membrane-damaging toxins produced by staphylococci. In Bacterial Toxins and Cell Membranes pp. 89–127 Jel-jaszewicz J., Wadström T. Edited by London: Academic Press;
    [Google Scholar]
  16. Reisfeld R.A., Lewis U.J., Williams D.E. 1962; Disk electrophoresis of basic proteins on polyacrylamide gels. Nature; London: 195281–283
    [Google Scholar]
  17. Rogolsky M. 1979; Nonenteric toxins of Staphylococcus aureus . Microbiological Reviews 43:320–360
    [Google Scholar]
  18. Smith G.M., Fitton J.E., Shaw W.V. 1979; Delta haemolysin of Staphylococcus aureus: purification, structure and biosynthesis. Society for General Microbiology Quarterly 6:147
    [Google Scholar]
  19. Spackman D.H., Stein W.H., Moore S. 1958; Automatic recording apparatus for use in the chromatography of amino acids. Analytical Chemistry 30:1190–1206
    [Google Scholar]
  20. Yoshida A. 1963; Staphylococcal δ-haemolysin. 1. Purification and chemical properties. Biochimica et biophysica acta 71:544–553
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-124-2-365
Loading
/content/journal/micro/10.1099/00221287-124-2-365
Loading

Data & Media loading...

Most cited Most Cited RSS feed