Functional activity of staphylococcal enterotoxin A requires interactions with both the alpha and beta chains of HLA-DR

James E. Dowd, Robert W. Karr, David R. Karp

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The staphylococcal enterotoxins, SEA and SEE, bind one zinc atom per molecule of protein. The presence of this metal atom enhances the binding of the toxins to MHC class II molecules, presumably through an interaction with histidine 81 of the β chain. L cell transfectants expressing HLA-DR1 and HLA-DR7 molecules, with mutations in either the α1 or β1 domains, were tested for their ability to bind SLA and present it to T cells. Cells expressing DR1 molecules with alanine at positions 77, 78, 80, 83, 84 and 85, or serine at position 79 could all bind SEA and present it to either polyclonal or monoclonal T cells. Most point mutations within the α-helical portion of the DR7 β chain had no effect on binding and presentation. However, substitution of histidine 81 with alanine, glutamate, or aspartate, abrogated SEA binding as well as T cell stimulation by the superantigen. This effect was also observed when the non-polymorphic aspartate, at position 76 was changed to alanine. Mutation of the asparagine at position 82 had an intermediate effect. Point mutations of the DR α chain had little effect on binding of SEA as determined by a flow cytometric assay. However, mutation of lysine at position 39 of the α chain and, to a lesser extent methionine at position 36, markedly decreased the ability of SEA to stimulate toxin-responsive mouse T cell hybridomas. Finally, the monoclonal antibody, L243 binds to the α chain of HLA-DR, and was able to block T cell activation by SEA without blocking SEA binding. These data support the model whereby HLA-DR has two binding sites for SEA. A low affinity site, present on the α chain, is required for T cell stimulation by the superantigen, but is insufficient to mediate toxin binding. High affinity binding of HLA-DR to SEA occurs solely through residues on the β chain, including both histidine 81 and aspartate 76.

Original languageEnglish (US)
Pages (from-to)1267-1274
Number of pages8
JournalMolecular Immunology
Volume33
Issue number16
DOIs
StatePublished - Nov 1996

Fingerprint

HLA-DR beta-Chains
HLA-DR alpha-Chains
T-Lymphocytes
HLA-DR Antigens
Histidine
Aspartic Acid
Alanine
Superantigens
Point Mutation
Mutation
HLA-DR7 Antigen
HLA-DR1 Antigen
Enterotoxins
Asparagine
Hybridomas
Methionine
Serine
Lysine
Staphylococcal enterotoxin A
Zinc

Keywords

  • HLA-DR
  • staphylococcal enterotoxin
  • superantigen

ASJC Scopus subject areas

  • Molecular Biology
  • Immunology

Cite this

Functional activity of staphylococcal enterotoxin A requires interactions with both the alpha and beta chains of HLA-DR. / Dowd, James E.; Karr, Robert W.; Karp, David R.

In: Molecular Immunology, Vol. 33, No. 16, 11.1996, p. 1267-1274.

Research output: Contribution to journalArticle

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abstract = "The staphylococcal enterotoxins, SEA and SEE, bind one zinc atom per molecule of protein. The presence of this metal atom enhances the binding of the toxins to MHC class II molecules, presumably through an interaction with histidine 81 of the β chain. L cell transfectants expressing HLA-DR1 and HLA-DR7 molecules, with mutations in either the α1 or β1 domains, were tested for their ability to bind SLA and present it to T cells. Cells expressing DR1 molecules with alanine at positions 77, 78, 80, 83, 84 and 85, or serine at position 79 could all bind SEA and present it to either polyclonal or monoclonal T cells. Most point mutations within the α-helical portion of the DR7 β chain had no effect on binding and presentation. However, substitution of histidine 81 with alanine, glutamate, or aspartate, abrogated SEA binding as well as T cell stimulation by the superantigen. This effect was also observed when the non-polymorphic aspartate, at position 76 was changed to alanine. Mutation of the asparagine at position 82 had an intermediate effect. Point mutations of the DR α chain had little effect on binding of SEA as determined by a flow cytometric assay. However, mutation of lysine at position 39 of the α chain and, to a lesser extent methionine at position 36, markedly decreased the ability of SEA to stimulate toxin-responsive mouse T cell hybridomas. Finally, the monoclonal antibody, L243 binds to the α chain of HLA-DR, and was able to block T cell activation by SEA without blocking SEA binding. These data support the model whereby HLA-DR has two binding sites for SEA. A low affinity site, present on the α chain, is required for T cell stimulation by the superantigen, but is insufficient to mediate toxin binding. High affinity binding of HLA-DR to SEA occurs solely through residues on the β chain, including both histidine 81 and aspartate 76.",
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AB - The staphylococcal enterotoxins, SEA and SEE, bind one zinc atom per molecule of protein. The presence of this metal atom enhances the binding of the toxins to MHC class II molecules, presumably through an interaction with histidine 81 of the β chain. L cell transfectants expressing HLA-DR1 and HLA-DR7 molecules, with mutations in either the α1 or β1 domains, were tested for their ability to bind SLA and present it to T cells. Cells expressing DR1 molecules with alanine at positions 77, 78, 80, 83, 84 and 85, or serine at position 79 could all bind SEA and present it to either polyclonal or monoclonal T cells. Most point mutations within the α-helical portion of the DR7 β chain had no effect on binding and presentation. However, substitution of histidine 81 with alanine, glutamate, or aspartate, abrogated SEA binding as well as T cell stimulation by the superantigen. This effect was also observed when the non-polymorphic aspartate, at position 76 was changed to alanine. Mutation of the asparagine at position 82 had an intermediate effect. Point mutations of the DR α chain had little effect on binding of SEA as determined by a flow cytometric assay. However, mutation of lysine at position 39 of the α chain and, to a lesser extent methionine at position 36, markedly decreased the ability of SEA to stimulate toxin-responsive mouse T cell hybridomas. Finally, the monoclonal antibody, L243 binds to the α chain of HLA-DR, and was able to block T cell activation by SEA without blocking SEA binding. These data support the model whereby HLA-DR has two binding sites for SEA. A low affinity site, present on the α chain, is required for T cell stimulation by the superantigen, but is insufficient to mediate toxin binding. High affinity binding of HLA-DR to SEA occurs solely through residues on the β chain, including both histidine 81 and aspartate 76.

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