Cooperative zinc binding in a staphylococcal enterotoxin A mutant mimics the SEA-MHC class II interaction

摘要

The structure of a mutant form of staphylococcal enterotoxin A (sEA) has been determined to 2.1 A resolution. The studied SEA substitution H187 → A187 (SEA_(H187A)) leads to an almost 10-fold reduction of the binding to major histocompatibility complex (MHC) class II. H187 is important for this interaction since it coordinates Zn~(2+). The zinc ion is thought to hold MHC class II and SEA together in a complex. Interestingly, only one of two molecules in the asymmetric unit binds Zn~(2+). H225, D227, a water molecule, and H44 from a symmetry-related molecule ligate Zn~(2+). The symmetry-related histidine is necessary for this substituted Zn~(2+) site to bind to Zn~(2+) at low zinc concentration (no Zn~(2+) added). Since a water molecule replaces the missing H187, H44 binds Zn~(2+) at the position where βH81 from MHC class II probably will bind. Dynamic light scattering analysis reveals that in solution as well as in the crystal lattice the SEA_(H187A) mutant forms aggregates. The substitution per se does not cause aggregation since wild-type SEA also forms aggregates. Addition of EDTA reduces the size of the aggregates, indicating a cross-linking function of Zn~(2+). In agreement with the biological function, the aggregation is weak (i.e. not revealed by gel filtration) and non-specific.