A mutational analysis of the binding of staphylococcal enterotoxins b and c3 to the T cell receptorb chain and MHC class II

LEDER L; LLERA ANDREA S; LAVOIE P; LEBEDEVA M; LI, HM; SEKALY RP; BOHACH G; GAHN PG; SCHLIEVERT, PM; KARJALAINEN, K; MARIUZZA, RA

JOURNAL OF EXPERIMENTAL MEDICINE

Año: 1998 vol. 187 p. 823 - 834

0022-1007

The three-dimensional structure of the complex between a T cell receptor (TCR)bchain (mouse Vb8.2Jb2.1Cb1) and the superantigen (SAG) staphylococcal enterotoxin C3 (SEC3) has been recently determined to 3.5 Å resolution. To evaluate the actual contribution of individual SAG residues to stabilizing the Vb–SEC3 complex, as well as to investigate the relationship between the affinity of SAGs for TCR and MHC and their ability to activate T cells, we measured the binding of a set of SEC3 and staphylococcal enterotoxin B (SEB) mutants to soluble recombinant TCRb chain and to the human MHC class II molecule HLA-DR1. Affinities were determined by sedimentation equilibrium and/or surface plasmon detection, while mitogenic potency was assessed using T cells from rearrangement-deficient TCR transgenic mice. We show that there is a clear and simple relationship between the affinity of SAGs for the TCR and their biological activity: the tighter the binding of a particular mutant of SEC3 or SEB to the TCRb chain, the greater its ability to stimulate T cells. We also find that there is an interplay between TCR–SAG and SAG–MHC interactions in determining mitogenic potency, such that a small increase in the affinity of a SAG for MHC can overcome a large decrease in the SAG’s affinity for the TCR. Finally, we observe that those SEC3 residues thatmake the greatest energetic contribution to stabilizing the Vb–SEC3 complex (“hot spot” residues) are strictly conserved among enterotoxins reactive with mouse Vb8.2, thereby providing a basis for understanding why SAGs having other residues at these positions show different Vb-binding specificities.