Isolation of a phosphoryl choline-binding protein from the hemolymph of the snail, Achatina fulica.
A phosphorylcholine-binding protein from the hemolymph of the snail Achatina fulica was purified to near homogeneity using a Sepharose phenylphosphorylcholine affinity column. The protein bound to the affinity column was eluted with 5 mM phosphorylcholine as a single symmetrical peak. The purified protein (400 Kda) contained 35-40% carbohydrate. On SDS-PAGE the protein separated into two bands of 20 and 24 Kda, and had a pI of 5.9. On immunodiffusion, antiserum to the snail phosphorylcholine binding protein did not cross-react against other phosphorylcholine binding proteins, like rat serum phosphorylcholine-binding protein (PCBP), limulus C-reactive protein (CRP), or human CRP. On pretreatment of the snail hemolymph with this antiserum, the hemagglutination titer of the hemolymph was markedly decreased. The purified snail phosphorylcholine binding protein agglutinated rabbit erythrocytes in the absence of divalent cation (Ca+2) but trace amount of Ca+2 increased its binding. The strongest inhibitor of the agglutination reaction was lactose, followed by melibiose and 2-deoxygalactose. The relationships of the snail phosphorylcholine binding protein to other hemolymph agglutinins and to CRPs are discussed in light of common phylogeny.
Mandal
C
C
Indian Institute of Chemical Biology, Jadavpur, Calcutta.
Biswas
M
M
Nagpurkar
A
A
Mookerjea
S
S
D000818:Animals; D002097:C-Reactive Protein / Q000276:immunology; D002241:Carbohydrates / Q000494:pharmacology; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000097:blood* / Q000276:immunology; D003429:Cross Reactions; D006023:Glycoproteins / Q000097:blood* / Q000276:immunology; D006386:Hemagglutination Tests; D006458:Hemolymph / Q000378:metabolism*; D006737:Horseshoe Crabs / Q000276:immunology; D006801:Humans; D005779:Immunodiffusion; D037102:Lectins / Q000276:immunology; D008970:Molecular Weight; D010767:Phosphorylcholine / Q000378:metabolism*; D012908:Snails / Q000276:immunology / Q000378:metabolism*; D013045:Species Specificity; D014714:Vertebrates / Q000276:immunology
D002241:Carbohydrates; D002352:Carrier Proteins; D006023:Glycoproteins; D037102:Lectins; C015323:limulin; D010767:Phosphorylcholine; D002097:C-Reactive Protein