Sialic acid-binding lectin with antibacterial activity from the horse mussel: further characterization and immunolocalization

Abstract

A heterogeneous sialic acid-binding lectin with affinity for bacterial LPS was isolated and partially characterized from hemolymph of the horse mussel Modiolus modiolus. [1]Using two-dimensional electrophoresis with immobilized pH gradients, the lectin revealed three subunits with different molecular weight and isoelectric points (pI); Mr14 (pI∼5.1 and ∼5.5), 17.5 (pI∼5.5) and 20 (pI∼4.9) kDa. The affinity purified lectin existed in its native state as aggregates, and by stepwise centrifugation it could be fractionated into molecular entities with distinct specificities towards human and/or horse erythrocytes (modiolin H and/or E activity, respectively). While the medium size entities (range ≤30 and <100 kDa) exhibited only modiolin E activity and the lowest size entities (range ≤5 and <10 kDa) demonstrated only modiolin H activity, the largest aggregates (≥100 kDa)expressed both activities. Antibacterial activity of the lectin has been observed against various marine bacteria, whereas the whole hemolymph was less effective. The lectin exhibited strong antibacterial effect against all tested strains of Vibrio anguillarum, Vibrio salmonicida, Vibrio viscosus, Vibrio wodanis, and Vibrio ordalii, slight effect on Aeromonas salmonicida salmonicida and Shewanella putrefaciens, and no inhibitory effect with Alteromonas sp. Hemolymph of the horse mussel demonstrated no antibacterial effect against A. salmonicida salmonicida, Alteromonas sp., Sh. putrefaciens and some strains of V. anguillarum, but slight effects against some strains of V. anguillarum and both strains of V. ordalii, and more predominantly against V. wodanis, V. salmonicida and V. viscosus. These results indicate that the lectin plays a role in elimination of bacteria. Circulating hemocytes were demonstrated to be the source of the lectins since granules of the hemocytes were immunoreactive to anti-hemolymph lectin antibody and protein A/gold labelling.