Backbone resonance assignment of Staphylococcal Enterotoxin H

Abstract

The staphylococcal enterotoxin H (SEH; 217 aa, 25 kD) belongs to a family of superantigens that cause a massive immune response upon simultaneous binding to the T cell receptor (TCR) and the major histocompatibility complex class II. The SEH-TCR interaction is weak and amenable to studies using NMR methodology. Essentially, 2 mg of U{(2)H, (13)C,(15)N}-labeled SEH was used for the complete sequential backbone assignment of SEH at 900 MHz. The protein secondary structure inferred from the chemical shift index (C(alpha) and C(beta)) is in very good agreement with the secondary structure elements of the X-ray structure.

The structure of superantigen complexed with TCR and MHC reveals novel insights into superantigenic T cell activation

“…The sequential assignment of the SEH protein was reported elsewhere 20 . According to the chemical shift index, the structural integrity of the SEH protein in solution shows no deviation from the reported crystal structures 16,20 . An NMR study of the transient interaction between SEH and the TCR was performed.…”

“…Unlabelled SEH was produced in E. coli K12 strain UL635 in line with methods published earlier 15 , and SEH labelled with NMR active isotopes were produced in line with methods described by Saline et al 20 Briefly, the protein was overexpressed in E. coli. Following isolation of the periplasmic protein fraction, SEH was subjected to cationic exchange chromatography (Resource S 6-ml column, GE Healthcare) on an ÄKTA explorer (GE Healthcare).…”

Molecular modelling and docking analysis of pleurocidin (an antimicrobial peptide) like peptides with enterotoxin H from Klebsilla pneumonia

“…They are associated with endophthalmitis and urinary tract infection (UTI) [4]. A detailed understanding of the molecular structure and function of Enterotoxin H is highly relevant [5-8]. Therefore, it is of interest to study the interaction of enterotoxin H with pleurocidin like peptides using molecular modelling (template PDB ID: 1YCE), Lig-Plot (ligand construction) and docking tools for therapeutic consideration.…”