Molecular structure of isolated MvspI, a variable surface protein of the fish pathogen
Adan, J.*; Yoshii, Shuhei*; Kono, Hidetoshi; Miyata, Makoto*
is a parasitic bacterium that causes necrosis in the gills of freshwater fishes. This study examines the molecular structure of its surface variable protein, MvspI, composed of 2002 amino acids. MvspI was isolated from Mycoplasma cells by a biochemical procedure to 92% homogeneity. Gel filtration and analytical ultracentrifugation suggested that this protein is a cylinder-shaped monomer with axes of 66 and 2.7 nm. Rotary shadowing transmission electron microscopy of MvspI showed that the molecule is composed of two rods 30 and 45 nm long; the latter rod occasionally features a bulge. The electron microscopy with a monoclonal antibody and epitope mapping showed that the bulge end of molecular image corresponds to the C-terminus of amino acid sequence. Partial digestion by various proteases suggested that the N-terminal part, comprised of 697 amino acids, is flexible. Analysis of the predicted amino acid sequence showed that the molecule features an N-terminal signal sequence and 16 repeats of about 90 residues; 15 positions exist between residues 88 and 1479, and the other position is between residues 1725 and 1807. The amino acid sequence of MvspI was assigned onto a molecular image obtained by electron microscopy, based on the features from both analyses. The present study is the first to elucidate the molecular shape of a surface variable protein of Mycoplasma.