Isolation, purification and characterization of beta-1,3-glucan binding protein from the plasma of marine mussel Perna viridis.
A beta-1,3-glucan binding protein (betaGBP) specific for laminarin (a beta-1,3-glucan) was detected for the first time in a mollusc, Perna viridis. betaGBP was isolated and purified from the plasma using laminarin precipitation and affinity chromatography on laminarin-Sepharose 6B, respectively. It agglutinated bakers yeast, bacteria, and erythrocytes and enhanced prophenoloxidase (proPO) activity of the plasma in a dose-dependent manner. The purified betaGBP appeared as a single band in native-PAGE and the purity was conformed by HPLC. The protein has a molecular weight estimate of 510kDa as determined by SDS-PAGE and in isoelectric focusing the purified betaGBP was focused as a single band at pI 5.3. betaGBP was found to possess inherent serine protease activity but lacked beta-1,3-glucanase activity and all these results suggest that plasma betaGBP of P. viridis functions as a recognition molecule for beta-1,3-glucan on the surface of microbial cell walls. This recognition and binding lead to the activation of the prophenoloxidase cascade mediated by the inherent serine protease activity of betaGBP. Presence of agglutinating activity and serine protease activity shows that betaGBP is a bifunctional protein. The findings are discussed in light of the importance of this protein in the innate immune response of P. viridis, and they implicate evolutionary link with similar proteins found in other invertebrates.
Jayaraj
S S
SS
Unit of Pathobiology, Department of Zoology, University of Madras, Guindy Campus, Chennai 600 025, India. jayaraj_ss73@yahoo.co.in
Thiagarajan
R
R
Arumugam
M
M
Mullainadhan
P
P
D000372:Agglutination Tests; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D006458:Hemolymph / Q000737:chemistry* / Q000276:immunology; D008024:Ligands; D014442:Monophenol Monooxygenase / Q000378:metabolism; D049883:Perna / Q000737:chemistry* / Q000276:immunology; D011485:Protein Binding; D012697:Serine Endopeptidases / Q000378:metabolism; D047071:beta-Glucans / Q000378:metabolism*
D002352:Carrier Proteins; D008024:Ligands; D047071:beta-Glucans; C033363:beta-1,3-glucan; D014442:Monophenol Monooxygenase; D012697:Serine Endopeptidases