Isolation and characterization of a hemagglutinin with affinity for lipopolysaccharides from plasma of the crayfish Pacifastacus leniusculus.
A hemagglutinin with a high specific activity against trypsinized rabbit erythrocytes was identified in plasma of the freshwater crayfish Pacifastacus leniusculus. The activity of this crayfish hemagglutinin could be inhibited by sialoglycoproteins such as porcine stomach mucin, bovine submaxillary mucin, fetuin, and ovalbumin. However, the involvement of sialic acid in its binding specificity could not be unambiguously proven. Furthermore, the hemagglutinating activity in the crayfish plasma could be specifically inhibited by lipopolysaccharide from E. coli K-235, which might indicate a recognition role for this hemagglutinin. This hemagglutinin, which accounts for less than 0.01% of the total plasma protein, was purified to near homogeneity using affinity chromatography on a Fetuin-Sepharose 4B column. The molecular mass of the unreduced protein as revealed by sodium dodecyl sulphate electrophoresis in polyacrylamide gel was found to be 420,000 Da. Upon reduction with dithiothreitol the hemagglutinin dissociated to several subunits with masses ranging from 65,000 to 80,000 Da. Affinoblotting with peroxidase labelled lectins indicated that the hemagglutinin was likely to be a glycoprotein.
Kopácek
P
P
Department of Physiological Botany, University of Uppsala, Sweden.
Grubhoffer
L
L
Söderhäll
K
K
D000818:Animals; D003400:Astacoidea / Q000737:chemistry*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000187:drug effects; D006023:Glycoproteins / Q000097:blood / Q000187:drug effects / Q000302:isolation & purification*; D006386:Hemagglutination Tests; D006388:Hemagglutinins / Q000097:blood / Q000187:drug effects / Q000302:isolation & purification*; D006458:Hemolymph / Q000737:chemistry*; D037102:Lectins / Q000097:blood / Q000187:drug effects / Q000302:isolation & purification*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008322:Mammals; D008970:Molecular Weight; D012795:Sialoglycoproteins / Q000494:pharmacology; D014357:Trypsin / Q000494:pharmacology
D006023:Glycoproteins; D006388:Hemagglutinins; D037102:Lectins; D008070:Lipopolysaccharides; D012795:Sialoglycoproteins; D014357:Trypsin