Protein A Isolated from <i>Staphylococcus aureus</i> after Digestion with Lysostaphin

Sjöquist, John; Meloun, B.; Hjelm, Hans

doi:10.1111/j.1432-1033.1972.tb02023.x

Cited by 288 publications

(107 citation statements)

References 36 publications

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“…The blocked N-terminus of the protein [8] indicates a short section to precede region D. If this is the case, it leads to the conclusion that the four regions D, A, B, and C most probably represent out-of-phase repetitive regions, i.e. regions overlapping the original ones.…”

Section: The Proposed Primary Structure Of the Total Fc-binding Portimentioning

confidence: 99%

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Structural Studies on the Four Repetitive Fc-Binding Regions in Protein A from Staphylococcus aureus

Sjödahl¹

1977

Eur J Biochem

121

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The covalent structures of the four highly homologous Fc-binding regions in protein A, regions D, A, B, and C, have been studied by enzymic fragmentations of previously isolated fragments originating from these regions and subsequent isolation of the generated peptides by ion-exchange chromatography, molecular-sieve chromatography, high-voltage paper electrophoresis and paper chromatography. The complete sequence of region B was elucidated by combining the results of Edman degradations on isolated fragment B peptides with the previously reported N-terminal sequence of the same fragment. Furthermore, Edman degradations of fragment D, A and C peptides differing from the region B sequence provided the structures of subregions not identical to corresponding subregions within region B. Thus, it is possible to propose a highly probable covalent structure for the N-terminal 27 000-molecular-weight portion of protein A responsible for the IgG-Fc-binding activities. However, it was not possible to assign the activities to specific structures within the regions.The sequence data indicate that not only mutual homology between the four regions exists, but also internal homologies within the regions. Furthermore, the data strongly supports the hypothesis of a stepwise gene fusion procedure being involved in the evolution of the protein.

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Section: The Proposed Primary Structure Of the Total Fc-binding Portimentioning

confidence: 99%

“…The protein is isolated by digestion with lysostaphin [8] and subsequent affinity chromatography on IgG-Sepharose [9]. One protein A molecule is able to bind two or more IgG molecules [8].…”

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confidence: 99%

Structural Studies on the Four Repetitive Fc-Binding Regions in Protein A from Staphylococcus aureus

Sjödahl¹

1977

Eur J Biochem

121

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“…aureus strain Cowan I (NCTC 8530) was grown as a batch culture in a stirred fermentor with a working volume of 100 1. The culture medium and the conditions for growth have been described [6]. The microorganisms were precultured on rotary shakers for 10-12 h at 37 "C in five 5-1 conical flasks each containing 1 1 of culture medium.…”

Section: Staphylococcus Aureus Preparationmentioning

confidence: 99%

“…Protein A, released from bacteria by digestion with lysostaphin [6], was purified by using affinity chromatography on IgG-Sepharose as described earlier [17].…”

Section: Prtparation Oj Protein Amentioning

confidence: 99%

Immunologically Active and Structurally Similar Fragments of Protein A from Staphylococcus aureus

Hjelm

Sjödahl

Sjöquist

1975

European Journal of Biochemistry

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To study the active site(s) in protein A, partial tryptic digestions of the protein and of intact Staphylococcus aureus were performed. Fragments which bind to the Fc‐part of human IgG were isolated by affinity chromatography on IgG‐Sepharose 4B and purified by ion‐exchange chromatography on phosphocellulose. From a partial tryptic digest of pure protein A at 30°C, pH 8.2 for 30 min we have isolated and characterized six active fragments with molecular weights ranging from 6000 to 8000. Two active fragments, obtained in high yields by digestion at pH 7.2 of intact protein‐A‐containing bacteria, were shown to be similar to two of the six characterized fragments from the digest of pure protein A. All fragments appeared to have similar amino acid sequences, judged by peptide mapping, specific staining and amino acid analysis; some are very possibly overlapping peptides. Each fragment probably contains only one active site region since all are monovalent in the Fc‐reaction when studied with a hemagglutination technique. The maximal molar yield of active fragments obtained from the digestion of pure protein A accounts for about 210% of the amount of protein A used. Thus protein A, suggested to consist of repeating units, should exhibit at least three similar if not identical active regions.

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“…Protein A reacts with the Fc-region of IgG from several species [3-51 and is able to bind two or more molecules of IgG per molecule [6] . This interaction has been utilized in methods for immunochemical and cell surface structure studies [7,8].…”

Section: Introductionmentioning

confidence: 99%

Repetitive sequences in protein a from Staphyloccus aureus: Three highly homologous Fc‐binding regions

Sjödahl¹

1976

FEBS Letters

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Protein A Isolated from Staphylococcus aureus after Digestion with Lysostaphin

Cited by 288 publications

References 36 publications

Structural Studies on the Four Repetitive Fc-Binding Regions in Protein A from Staphylococcus aureus

Structural Studies on the Four Repetitive Fc-Binding Regions in Protein A from Staphylococcus aureus

Immunologically Active and Structurally Similar Fragments of Protein A from Staphylococcus aureus

Repetitive sequences in protein a from Staphyloccus aureus: Three highly homologous Fc‐binding regions

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