Purification and some chemical and physical properties of staphylococcal enterotoxin A

Schantz, Edward J.; Roessler, William G.; Woodburn, Margy; Lynch, Joseph M.; Jacoby, Henry M.; Silverman, Sidney J.; Gorman, Joseph C.; Spero, Leonard

doi:10.1021/bi00753a009

Cited by 92 publications

(55 citation statements)

References 24 publications

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“…Our data agree with their sequence data, except that we predict an extra Tyr residue at position 88 and we were able to order residues 99 through 103. The C-terminal residue is predicted to be a Ser residue, which agrees with protein analysis (42).…”

Section: Resultssupporting

confidence: 78%

Nucleotide sequence of the type A staphylococcal enterotoxin gene

1988

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We determined the nucleotide sequence of the gene encoding staphylococcal enterotoxin A (entA). The gene, composed of 771 base pairs, encodes an enterotoxin A precursor of 257 amino acid residues. A 24-residue N-terminal hydrophobic leader sequence is apparently processed, yielding the mature form of staphylococcal enterotoxin A (Mr, 27,100). Mature enterotoxin A has 82, 72, 74, and 34 amino acid residues in common with staphylococcal enterotoxins B and Cl, type A streptococcal exotoxin, and toxic shock syndrome toxin 1, respectively. This level of homology was determined to be significant based on the results of computer analysis and biological considerations. DNA sequence homology between the entA gene and genes encoding other types of staphylococcal enterotoxins was examined by DNA-DNA hybridization analysis with probes derived from the entA gene. A 624-base-pair DNA probe that represented an internal fragment of the entA gene hybridized well to DNA isolated from EntE+ strains and some EntA+ strains. In contrast, a 17-base oligonucleotide probe that encoded a peptide conserved among staphylococcal enterotoxins A, B, and Cl hybridized well to DNA isolated from EntA', EntB+, EntC1+, and EntD+ strains. These hybridization results indicate that considerable sequence divergence has occurred within this family of exotoxins.

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Section: Resultssupporting

confidence: 78%

Nucleotide sequence of the type A staphylococcal enterotoxin gene

1988

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“…Staphylococcal enterotoxin A (SEA), one of the 21 S. aureus toxins identified to date, is the most commonly encountered biotoxin involved in staphylococcal food poisoning outbreaks [1]. SEA is a small monomeric protein (MW = 28 kDa) with high thermal and proteolytic stability [2] so that even consumption of cooked food can be deleterious to health if contaminated.…”

Section: Introductionmentioning

confidence: 99%

Gold colloid-nanostructured surfaces for enhanced piezoelectric immunosensing of staphylococcal enterotoxin A

Haddada

Salmain

Boujday

2018

Sensors and Actuators B: Chemical

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Highlights Gold-and silicon-coated quartz sensors were nanostructured with gold nanoparticles  A capture layer comprising an anti-SEA antibody was built up on the sensor surface  Nanostructuration improved sensor response owing to better accessibility of antibodies  With a sandwich-type assay, the limit of detection was 1 ng SEA/mL in 25 min AbstractWe describe the use of gold nanoparticles (AuNP) as a nanostructuring agent on quartz crystal sensor chips to engineer staphylococcal enterotoxin A (SEA) piezoelectric biosensors with amplified response. AuNPs were assembled on gold-or silicon-coated quartz crystal sensor chips by a wet chemistry process involving their chemisorption to preformed thiol and amine terminated Self-Assembled Monolayers (SAMs). The purpose of this nanostructuration was to modify the topography of the surface and improve the accessibility of the binding sites on the surface of the sensor chips. Biointerfaces, comprising a polyclonal antibody against staphylococcal enterotoxin A (SEA), were further built up on these gold nanoparticle-coated sensors and their ability to capture SEA was monitored in real time with a quartz crystal microbalance with dissipation monitoring. It was found out that, although the surface density in capture antibody was similar on both nanostructured and planar sensors, the sensor response, expressed as frequency shift recorded during the binding of SEA to the antibody, was significantly higher for the nanostructured sensors as compared to the planar ones. All the same, the limit of detection was lower for the nanostructured sensors: 8 ng/mL vs 20 ng/mL for the planar sensors. This was rationalized by a possibly better accessibility of the antigen binding sites rather than a consequence of specific surface increase. Using a sandwich type assay, gold nanoparticles coated silicon quartz sensor chips provided the lowest limit of detection of ca. 1 ng/mL in a total assay time of 25 min. 2

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“…Exceptions to this rule have been documented by us and others. These are proteins that stimulate T cells bearing the appropriate V13 element almost independently ofcontributions from other parts of the TCRs borne by the T cells (9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22). Such proteins, called superantigens because they stimulate large numbers ofT cells, are encoded by microorganisms including retroviruses, bacteria, and mycoplasma (22)(23)(24)(25)(26)(27)(28)(29).…”

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confidence: 99%

A method for production of antibodies to human T-cell receptor beta-chain variable regions.

Choi

Kotzin

Lafferty

et al. 1991

Proc. Natl. Acad. Sci. U.S.A.

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Mouse T-cell hybridomas bearing human Vp elements were produced by transfection of human/mouse hybrid T-cell receptor 18-chain genes into a mouse T-cell hybridoma lacking an endogenous #-chain gene. These hybridomas were entirely mouse in origin except for the human Vp region. These cells were used to immunize mice against human V13 elements. Mouse monoclonal antibodies have thus been generated against human Vp13.1 and -13.2. We expect that the method outlined in this paper will be useful in the production of monoclonal antibodies specific for other human V13 or V.elements.The receptor for antigen on most peripheral T cells is made of a and 13 chains. Usually these T-cell receptors (TCRs) engage antigen in the form of peptides, derived from the antigen, bound to cell surface major histocompatibility complex (MHC) molecules (1-4). The specificity of the TCR for this complex ligand is determined by the five variable elements (Va, Ja, V13, Do, and JO3) of the a and 13 chains as well as non-germ-line-encoded junctional regions (5-8). Exceptions to this rule have been documented by us and others. These are proteins that stimulate T cells bearing the appropriate V13 element almost independently ofcontributions from other parts of the TCRs borne by the T cells (9-22). Such proteins, called superantigens because they stimulate large numbers ofT cells, are encoded by microorganisms including retroviruses, bacteria, and mycoplasma (22-29).There is considerable interest in the control and consequences of the T-cell repertoire in humans and other species. Obviously a detailed understanding of the T-cell repertoire would require knowledge of all the V,, Ja, V13, D13, and Jo combinations present in a given individual. Such knowledge will not be available in the foreseeable future. However, much has already been learned from studies of the distribution between animals of just one of the TCR variable elements, V13. Examination of V13 expression in mice has been used to study positive selection, T-cell tolerance, autoimmunity, and the consequences of attack by superantigens (22).Similarly, V13 expression in humans has been used to study the effects of superantigens and also to examine the repertoires of T cells involved in inflammatory and autoimmune disease (18,19,29,30 MATERIALS AND METHODSPreparation of RNA and cDNA and Amplification by QPCR. Total RNA was prepared from phytohemagglutinin-, antihV,013

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Purification and some chemical and physical properties of staphylococcal enterotoxin A

Cited by 92 publications

References 24 publications

Nucleotide sequence of the type A staphylococcal enterotoxin gene

Nucleotide sequence of the type A staphylococcal enterotoxin gene

Gold colloid-nanostructured surfaces for enhanced piezoelectric immunosensing of staphylococcal enterotoxin A

A method for production of antibodies to human T-cell receptor beta-chain variable regions.

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