Immunological analysis of chemoattractive proteins from earthworm to garter snakes.
1. Two sets of polyclonal antibodies to two highly purified prey-derived snake-attractive proteins, a low molecular weight (3000) protein and a 20,000 mol. wt protein, were generated in rabbits. 2. They are immunospecific for their respective purified immunogens and do not cross-react with each other. 3. Eight prey-derived proteins that elicit attack by garter snakes (Thamnophis sirtalis) from earthworms (Lumbricus terrestris) were analyzed with these antibodies, and can be assigned to three distinct groups on the basis of their antigenic properties. 4. Unfolding or denaturation of the low molecular weight protein did not alter its antigenic activity to its polyclonal antibodies, suggesting the antigenic epitopes contain contiguous amino acid sequences. 5. In contrast, unfolding of the 20,000 mol. wt protein resulted in a loss of its binding with antibodies, suggesting that the epitope of this protein contains noncontiguous amino acid sequences. 6. The snake-attractivity of the 20,000 mol. wt protein could not be neutralized by reacting it with its antiserum, suggesting that the antigenic determinant (the epitope) of the antigen is not an integral part of the attractive domain of the ES20 protein. 7. In contrast, the attractivity of the purified low molecular weight protein could be neutralized by the polyclonal antibodies.
Wang
D
D
Department of Biochemistry, SUNY Health Science Center, Brooklyn 11203.
Chen
P
P
Halpern
M
M
D000818:Animals; D000906:Antibodies / Q000502:physiology; D000918:Antibody Specificity / Q000502:physiology; D000941:Antigens / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D002630:Chemotactic Factors / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D004558:Electric Stimulation; D015151:Immunoblotting; D005779:Immunodiffusion; D009835:Oligochaeta / Q000276:immunology* / Q000502:physiology; D011489:Protein Denaturation; D012911:Snakes / Q000276:immunology* / Q000502:physiology; D013438:Sulfhydryl Compounds / Q000302:isolation & purification
D000906:Antibodies; D000941:Antigens; D002630:Chemotactic Factors; D013438:Sulfhydryl Compounds