Item

Abstract

The solution structure of histidine-containing phosphocarrier protein from Staphylococcus carnosus was determined by two- and three-dimensional nuclear magnetic resonance (NMR) spectroscopy on uniformly 15N-enriched protein. The main structural element is an antiparallel beta-pleated sheet with four strands A, B, C, and D arranged with the topology A-D-B-C. Strand A comprises residues 2 to 8, strand B residues 32 to 37, strand C reidues 40 to 43, and strand D residues 59 to 66. Three right-handed helices are arranged on top of the beta-pleated sheet. Helix a reaches from residue 16 to 29, helix b from residue 48 to 53, and helix c from residue 72 to 83. Strands B and C of the beta-pleated sheet are connected by a type II turn. The hydroxyl proton of Ser-31 is ex-changing with the solvent so slowly that cross peaks can be detected in two-dimensional NMR spectra based on homonuclear J-couplings. The imidazole ring of the active-center His-15, which is partly charged in the structure determined at pH 7.2, is located above the N-terminal end of helix a, perpendicular to its axis. The Nδ1 atom of His-15, accepting the phosphoryl from enzyme I, is exposed to the solvent