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Chemical differences in the venoms of genetically different snakes
Abstracts
299
Tu, A. T., TOOM, P. M. and MURDOCK, D. S. (Department Chemistry; Interdepartmental Curriculum in Toxicology, Utah State University, Logan, Utah). Chemical differences in the venoms of genetically different snakes. TWENTY-SEVENdipeptides
and four tripeptides were used for determining the site of peptide hydrolysis by twelve rattlesnake venoms (Genera: Crotalm and Sistrarus). Peptides not hydroIysed by any of the rattlesnake venoms were Gly-~-Asp, Gly-L-Glu, L-Leu-L-Phe, &Leu-DL-Val, L-Lys-Gly, and L-Phe-L-Phe. Each of the triueutides was hvdrolvsed at both _ De&de linkages. - Lwith the linkage containing the N-terminal - _ _ amino acid being hydrolysed first. Of the twelve rattlesnake venoms investigated, there were no differences in substrate specificities resulting from either geographical origin or species differences. The actions of eight cobra venoms (Genera: Naja and Ophiophagus) were studied on twenty-four dipeptides and five tripeptides. The peptides Gly-~-Asp, Gly-L-Glu, L-Lys-Gly, and DL-Leu-DL-Val were not hydrolysed by any of the venoms. Three dipeptides, Gly-r_-Thr, L-Leu-L-Phe, and L-Leu-L-Tyr, were split by some of the cobra venoms investigated. As was the case with the rattlesnake venoms, the peptide bond of the tripeptide adjacent to the N-terminal amino acid was hydrolysed first. Regardless of the geographical distribution, the cobra venoms exhibited quite similar substrate specificities. However, the venoms of 0. I?annah and N. nuja atra appeared td be slightly different than the venoms of the other cobras. Among all the dipeptides investigated, the onIy difference iti substrate specificity between the rattleWhile six of the eight cobra venoms snake and the cobra venoms was detected on the peptide L-Leu-L-Phe. investigated hydrolysecl this peptide, none of the thirteen rattlesnake venoms studied split L-Leu-L-Phe. Of the sixty-eight venoms investigated (37 Crotalidae, 8 Viperidae, 21 Elapidae and 2 Hydrophiidae), none of! the Elapidae and Hydroohiidae’venoms, with the exception of 0. hannah, hydrolysed TAME and BAEE. In”gen&ral Viperidae venoms ‘gave lower $roteinase activities than those of Crotalidae using these substrates. It appears that there is no correlation between proteinase activities and the genera of poisonous snakes, as’ there is a large variation within each genus.
,WATT, D. D. and MCINTOSH,M. (Division Biological Sciences, Midwest Research
Institute, Kansas City, Missouri; Department of Microbiology, Kansas University Medical Center, Kansas City, Kansas). Molecular aspects of neurotoxic principle in venom of the scorpion Centruroides sculpturatus.
hIDIRECTevidence is presented
that the toxicity of the venom of Centruroides sc&turatus may be associated with the formation of acetylchoIine. This evidence was not supported in potentiation and inhibition studies using chemicals known to affect the acetylcholine transmitter system. From these studies, we feel quite sure that the toxic moiety is not associated with acetylcholine destruction,
SLOTTA, K. H. (Department
Biochemistry, Medical Research Division, Miami, Florida). The direct and indirect hemolytic factors from animal venoms.
THE ACTIVITIES of the two lytic factors in snake and bee venoms, phospholipase A and the direct lytic factor (DLF), were compared. Whereas these factors constitute two-thirds of bee venom, they are present in snake venoms in only very small amounts. Various methods (electrophoresis on paper, starch gel or other carriers) are available for the separation of the protein-polypeptide mixtures from crude snake venoms and for the isolation of the two lytic factors. However, the yields are usually poor and a new approach seemed to be indicated. Elapidae venoms from N. naja and Hemachatus haemachatus were subjected to gel titration and ion exchange chromatography. Although these two snakes belong to the same family, we found considerable diiYerences in their venoms. As the polypeptides of N. haemac/zatus vary in their molecular weights, they could easily be separated into three groups by gel filtration. This was not the case with N. naja venom where the individual components are distinguished primarily by their ele&ric charge. Thus, the first separation had to be carried out on ion exchange columns. Electrophoresis on cellulose polyacetate strips proved to be an excellent analytical tool for the detection . of the components in al1 fractions. Even minute quantities of DLF in the eluates from the columns could be found by hemolysis of washed red cells when a few pg of pure bee phospholipase A were added to the test solutions.
299
Tu, A. T., TOOM, P. M. and MURDOCK, D. S. (Department Chemistry; Interdepartmental Curriculum in Toxicology, Utah State University, Logan, Utah). Chemical differences in the venoms of genetically different snakes. TWENTY-SEVENdipeptides
and four tripeptides were used for determining the site of peptide hydrolysis by twelve rattlesnake venoms (Genera: Crotalm and Sistrarus). Peptides not hydroIysed by any of the rattlesnake venoms were Gly-~-Asp, Gly-L-Glu, L-Leu-L-Phe, &Leu-DL-Val, L-Lys-Gly, and L-Phe-L-Phe. Each of the triueutides was hvdrolvsed at both _ De&de linkages. - Lwith the linkage containing the N-terminal - _ _ amino acid being hydrolysed first. Of the twelve rattlesnake venoms investigated, there were no differences in substrate specificities resulting from either geographical origin or species differences. The actions of eight cobra venoms (Genera: Naja and Ophiophagus) were studied on twenty-four dipeptides and five tripeptides. The peptides Gly-~-Asp, Gly-L-Glu, L-Lys-Gly, and DL-Leu-DL-Val were not hydrolysed by any of the venoms. Three dipeptides, Gly-r_-Thr, L-Leu-L-Phe, and L-Leu-L-Tyr, were split by some of the cobra venoms investigated. As was the case with the rattlesnake venoms, the peptide bond of the tripeptide adjacent to the N-terminal amino acid was hydrolysed first. Regardless of the geographical distribution, the cobra venoms exhibited quite similar substrate specificities. However, the venoms of 0. I?annah and N. nuja atra appeared td be slightly different than the venoms of the other cobras. Among all the dipeptides investigated, the onIy difference iti substrate specificity between the rattleWhile six of the eight cobra venoms snake and the cobra venoms was detected on the peptide L-Leu-L-Phe. investigated hydrolysecl this peptide, none of the thirteen rattlesnake venoms studied split L-Leu-L-Phe. Of the sixty-eight venoms investigated (37 Crotalidae, 8 Viperidae, 21 Elapidae and 2 Hydrophiidae), none of! the Elapidae and Hydroohiidae’venoms, with the exception of 0. hannah, hydrolysed TAME and BAEE. In”gen&ral Viperidae venoms ‘gave lower $roteinase activities than those of Crotalidae using these substrates. It appears that there is no correlation between proteinase activities and the genera of poisonous snakes, as’ there is a large variation within each genus.
,WATT, D. D. and MCINTOSH,M. (Division Biological Sciences, Midwest Research
Institute, Kansas City, Missouri; Department of Microbiology, Kansas University Medical Center, Kansas City, Kansas). Molecular aspects of neurotoxic principle in venom of the scorpion Centruroides sculpturatus.
hIDIRECTevidence is presented
that the toxicity of the venom of Centruroides sc&turatus may be associated with the formation of acetylchoIine. This evidence was not supported in potentiation and inhibition studies using chemicals known to affect the acetylcholine transmitter system. From these studies, we feel quite sure that the toxic moiety is not associated with acetylcholine destruction,
SLOTTA, K. H. (Department
Biochemistry, Medical Research Division, Miami, Florida). The direct and indirect hemolytic factors from animal venoms.
THE ACTIVITIES of the two lytic factors in snake and bee venoms, phospholipase A and the direct lytic factor (DLF), were compared. Whereas these factors constitute two-thirds of bee venom, they are present in snake venoms in only very small amounts. Various methods (electrophoresis on paper, starch gel or other carriers) are available for the separation of the protein-polypeptide mixtures from crude snake venoms and for the isolation of the two lytic factors. However, the yields are usually poor and a new approach seemed to be indicated. Elapidae venoms from N. naja and Hemachatus haemachatus were subjected to gel titration and ion exchange chromatography. Although these two snakes belong to the same family, we found considerable diiYerences in their venoms. As the polypeptides of N. haemac/zatus vary in their molecular weights, they could easily be separated into three groups by gel filtration. This was not the case with N. naja venom where the individual components are distinguished primarily by their ele&ric charge. Thus, the first separation had to be carried out on ion exchange columns. Electrophoresis on cellulose polyacetate strips proved to be an excellent analytical tool for the detection . of the components in al1 fractions. Even minute quantities of DLF in the eluates from the columns could be found by hemolysis of washed red cells when a few pg of pure bee phospholipase A were added to the test solutions.