CRYSTAL-STRUCTURE OF A BACTERIAL SIALIDASE (FROM SALMONELLA-TYPHIMURIUM LT2) SHOWS THE SAME FOLD AS AN INFLUENZA-VIRUS NEURAMINIDASE

Sialidases (EC 3.2.1.18 or neuraminidases) remove sialic acid from sia loglycoconjugates, are widely distributed in nature, and have been imp licated in the pathogenesis of many diseases. The three-dimensional st ructure of influenza virus sialidase is known, and we now report the t hree-dimensional structure of a bacterial sialidase, from Salmonella t yphimurium LT2, at 2.0-angstrom resolution and the structure of its co mplex with the inhibitor 2-deoxy-2,3-dehydro-N-acetylneuraminic acid a t 2.2-angstrom resolution. The viral enzyme is a tetramer; the bacteri al enzyme, a monomer. Although the monomers are of similar size (almos t-equal-to 380 residues), the sequence similarity is low (almost-equal -to 15%). The viral enzyme contains at least eight disulflde bridges, conserved in all strains, and binds Ca2+, which enhances activity; the bacterial enzyme contains one disulfide and does not bind Ca2+. Compa rison of the two structures shows a remarkable similarity both in the general fold and in the spatial arrangement of the catalytic residues. However, an rms fit of 3.1 angstrom between 264 C(alpha) atoms of the S. typhimurium enzyme and those from an influenza A virus reflects so me major differences in the fold. In common with the viral enzyme, the bacterial enzyme active site consists of an arginine triad, a hydroph obic pocket, and a key tyrosine and glutamic acid, but differences in the interactions with the O4 and glycerol groups of the inhibitor refl ect differing kinetics and substrate preferences of the two enzymes. T he repeating ''Asp-box'' motifs observed among the nonviral sialidase sequences occur at topologically equivalent positions on the outside o f the structure. Implications of the structure for the catalytic mecha nism, evolution, and secretion of the enzyme are discussed.