The amino acid sequence of the delta haemolysin of <i>Staphylococcus aureus</i>

Fitton, John E.; Dell, Anne; Shaw, William V.

doi:10.1016/0014-5793(80)81170-7

Cited by 136 publications

(80 citation statements)

References 22 publications

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“…Sequencing data firmly in agreement with the sequence found by Fitton et al (1980) also suggest that the delta haemolysin is the major component. However, it is believed that perha& a family of similar molecules are purified by the methods employed including the two forms of delta haemolysin.…”

Section: Discussionsupporting

confidence: 85%

“…Lysine residues and the amino-terminal residue are not usually seen due to the method of attachment of the polypeptides to the glass support. This sequence, together with the amino acid analyses of the products, suggested that their amino acid sequences were identical with that found by Fitton et al (1980) for delta haemolysin purified by Dr N. G. Heatley by his method from S. aureus 186X, containing N-terminal methionine. Repeated attempts to determine the N-terminal sequence of delta haemolysin purified by the method of Kreger et al (1971) were unsuccessful, suggesting that the terminal amino acid was blocked.…”

Section: N-terminal Amino Acid Sequence Analysissupporting

confidence: 77%

“…Additional bands such as were noted previously were again observed in the delta haemolysin purified by the alumina or hydroxylapatite methods. Insulin B chain and melittin, which have molecular weights very similar to that calculated for delta haemolysin from amino acid sequence analysis (Fitton et al, 1980), also ran as diffuse bands under the same conditions (results not shown). This observation suggests that, with the conditions employed for electrophoresis under denaturing conditions, the diffuseness of the delta haemolysin band is a simple consequence of molecular size.…”

Section: Polyacrylamide Gel Electrophoresismentioning

confidence: 53%

“…Repeated attempts to determine the N-terminal sequence of delta haemolysin purified by the method of Kreger et al (1971) were unsuccessful, suggesting that the terminal amino acid was blocked. Mass spectrometry of delta haemolysin purified by the method of Heatley (1971) from S, aureus 186X showed that a proportion of it contained N-formylmethionine (Fitton et al, 1980). When the Kreger preparation was treated with 1 M-HCl in methanol for 1 h at room temperature to unblock the amino-terminus (Fitton et al, 1980), subsequent sequence analysis revealed that the first ten residues were the same as in the other preparations.…”

Section: N-terminal Amino Acid Sequence Analysismentioning

confidence: 99%

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Comparison of Three Methods for the Purification of the Delta Haemolysin of Staphylococcus aureus

Smith¹,

Shaw²

1981

Microbiology

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Three methods for the purification of Staphylococcus aureus delta haemolysin were compared (Kantor et al., 1972; Kreger et al., 197 1 ; Heatley, 197 1). 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 (197 1) represented not less than 70% of the product.

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Section: Discussionsupporting

confidence: 85%

Section: N-terminal Amino Acid Sequence Analysissupporting

confidence: 77%

Section: Polyacrylamide Gel Electrophoresismentioning

confidence: 53%

Section: N-terminal Amino Acid Sequence Analysismentioning

confidence: 99%

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Comparison of Three Methods for the Purification of the Delta Haemolysin of Staphylococcus aureus

Smith¹,

Shaw²

1981

Microbiology

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“…Although the delta toxin monomer consists of a 26 amino acid residue polypeptide [5], the gel filtration results reported in the present study show that the toxin is present as a tetramer in aqueous solution at extremes of pH and in aqueous alcoholic solution. The observation that the toxin is also tetrameric in buffered 6 M guanidine-HCl indicates that this form of the toxin is extremely stable in aqueous solution, indeed, monomeric toxin has not been observed under any aqueous solvent conditions employed to date, although monomers may be present in solvents of lower dielectric constant.…”

Section: Discussioncontrasting

confidence: 49%