Staphylococcal enterotoxin A: Partial unfolding caused by high pressure or denaturing agents enhances superantigenicity.
The effect of transient exposure of Staphylococcus aureus enterotoxin A (SEA) to high pressure and/or denaturing agents was examined by assessing the toxin superantigenicity and immunoreactivity, and by monitoring pressure-induced changes in fluorescence emission spectra. Pressurization of SEA at 600 MPa and 45 degrees C in Tris-HCl buffer (20 mM, pH 7.4) resulted in a marked increase in both T-cell proliferation (superantigenicity) and immunoreactivity. In opposite, pressurization at 20 degrees C did not change significantly SEA superantigenicity and immunoreactivity, indicating some toxin baro-resistance. Exposure of SEA to 8 M urea at atmospheric pressure or at 600 MPa and 20 degrees C, also led to a marked increase of superantigenicity (but not of immunoreactivity). In contrast, exposure of SEA to sodium-dodecylsulfate (30 mM) led to an increase of immunoreactivity with some effect on superantigenicity after pressurization at 45 degrees C only. High pressure up to 600 MPa induced spectral changes which at 20 degrees C were fully reversible upon decompression. At 45 degrees C, however, a sharp break of the centre of spectral mass mainly due to tryptophan residues was observed at 300 MPa, and irreversible spectral changes mainly related to tyrosine residues subsisted after pressure release, indicating a marked protein conformational transition. Urea 8 M further increased SEA structural changes at 600 MPa and 20 degrees C. These results indicate that SEA, under a combination of high pressure and mild temperature, as well as in the presence of urea, partly unfolds to a structure of strongly increased T-cell proliferative ability.
René-Trouillefou
Malika
M
Université Montpellier 2, UMR Ingénierie des Agropolymères et Technologies Emergentes, Equipe de Biochimie et Technologie Alimentaires, F-34095 Montpellier, France.
Benzaria
Amal
A
Marchal
Stéphane
S
Lange
Reinhard
R
Caporiccio
Bertrand
B
Dumay
Eliane
E
D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects*; D004768:Enterotoxins / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D006358:Hot Temperature; D011312:Pressure; D011489:Protein Denaturation; D017510:Protein Folding; D051381:Rats; D017207:Rats, Sprague-Dawley; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013211:Staphylococcus aureus; D013501:Surface-Active Agents / Q000737:chemistry; D013601:T-Lymphocytes / Q000276:immunology*; D014508:Urea / Q000737:chemistry*
D004768:Enterotoxins; D013501:Surface-Active Agents; D012967:Sodium Dodecyl Sulfate; C026066:enterotoxin A, Staphylococcal; D014508:Urea