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Isolation and characterization of a non-heme iron-containing glycopeptide from the venom of Vipera russelli
Abstracts of Papers
527
P. J. DEORAs and N. E. VAD. (Haffkine Institute, Bombay-12). Lung reduction in snakes and toxicity of salivary secretions. The salivary secretions of Dryophis nasutus, a snake belonging to Colubridae, were mildly toxic to mice . Similar secretions from Ptyas mucosus caused little distress, and those from Natrix piscator were non-toxic. Both these snakes also belong to Colubridae . Gross anatomy of the glands indicated that some serrous cells were visible in Dryophis nasutus, while the rest had only mucus cells. Detailed anatomical studies on glands of some snakes belonging to Typhlopidae, Boidae, Colubridae, Elapidae and Viperidae were performed and the toxicity tested . During these studiesattention was also given to other important anatomical differences. It was noted that in some Colubridae there was reduction in the size of one lung lobe, while in Elapidae and Viperidae one lobe was completely absent . Lung lobes were studied for Resazurin reaction, protein content and dehydrogenase activity. The last activity in nonpoisonous snakes was 0-34 units/mg protein and in poisonous snakes it was 4-6 . This activity in Dryophis nasutus was 5-4 and it is supposed to be a non-poisonous snake. Bubbling of oxygen gas through the concentrated venom solution of Bungarus Caeruleus for fifteen min, decreased the toxicity from 1/80,000 to 1/12,000. The studies indicate a correlation between predominance of serrous cells in the gland with the reduction or atrophy of one lung lobe . P. J . DEORAS and N. E. VAD. (Haffkine Institute, Bombay-12) . Vacuum method of milking
and significance of venom out-put data.
Extraction of venom by a vacuum method was found helpful in the case of Bungarus caeruleus. The venom out-put from the same species of snake from different geographical regions was studied for toxicity . Snake bite data for different locations on the body and at different seasons were scrutinized. This was correlated with the differential output in venom in different months . The utility and significance of such data are discussed. G. D. DumTtzov . (Department of Organic Chemistry, Institute of Chemical Technology, Sofia 56, Bulgaria). Isolation and characterization of a non-heme iron-containing glycopeptide from the venom of Vipera russelli. A glyeopeptide containing non-heme iron has been isolated from the venom of the snake Vipera russelli. The purification procedure includes four steps: starch block electrophoresis, gel filtration on Sephadex G-75, ion exchange chromatography on QÀE-Sephadex and dialysis through Visking Co . cellophane tubings, 8/32 . Homogeneity was verified by acrylamide disc electrophoresis and N-terminal amino acid determination . The N-terminal group was found to be alanine. Amino acid analysis and spectral characterization were performed. In the carbohydrate portion after acidic hydrolysis were identified glucose, glucosamine and silaic acid . It was demonstrated by gel filtration under different conditions that this component formed complexes with some high molecular proteins of the venom, being in a state of associationdissociation equilibrium . The glycopeptide revealed a biological activity when tested on mice . This is the first time that such a substance was found in snake venom. M. Dzouic and I. L. BoNTA. (Department of Pharmacology, Medical Faculty Rotterdam, Rotterdam, The Netherlands) . Effect of Naja naja venom and a heparin-precipitable portion on threshold to nocioceptive and convulsive stimuli on rats . Observations concerning the analgesic and anti-epileptic properties of cobra venom were reported in the past but contradictory data in man and laboratory animals were also published (Lusn?vnv and WAD, 1954 ; Wnjt"m, 1960 ; VoRoBEv et al., 1968 ; D'AmouR and SMuTx, 1941). A reappraisal of the question on animal experiments was the aim of the present work. We used crude Naja ngla venom and two portions of it obtained by addition of heparin to the venom solution, and subsequent centrifugation. The two materials thus obtained were the supernatant containing the less basic and the precipitate containing the more basic components of the venom. Heparin was removed by addition of protamine sulphate and subsequent centrifugation . Rats were used throughout and the venom or its portions were administered s .c . As judged by the LD . the supernatant, containing the neurotoxin, was slightly more toxic (0295 mg/kg) and the precipitate considerably less toxic (3-10 mg/kg) than the full venom TOXICON 1972 VoL 10.
527
P. J. DEORAs and N. E. VAD. (Haffkine Institute, Bombay-12). Lung reduction in snakes and toxicity of salivary secretions. The salivary secretions of Dryophis nasutus, a snake belonging to Colubridae, were mildly toxic to mice . Similar secretions from Ptyas mucosus caused little distress, and those from Natrix piscator were non-toxic. Both these snakes also belong to Colubridae . Gross anatomy of the glands indicated that some serrous cells were visible in Dryophis nasutus, while the rest had only mucus cells. Detailed anatomical studies on glands of some snakes belonging to Typhlopidae, Boidae, Colubridae, Elapidae and Viperidae were performed and the toxicity tested . During these studiesattention was also given to other important anatomical differences. It was noted that in some Colubridae there was reduction in the size of one lung lobe, while in Elapidae and Viperidae one lobe was completely absent . Lung lobes were studied for Resazurin reaction, protein content and dehydrogenase activity. The last activity in nonpoisonous snakes was 0-34 units/mg protein and in poisonous snakes it was 4-6 . This activity in Dryophis nasutus was 5-4 and it is supposed to be a non-poisonous snake. Bubbling of oxygen gas through the concentrated venom solution of Bungarus Caeruleus for fifteen min, decreased the toxicity from 1/80,000 to 1/12,000. The studies indicate a correlation between predominance of serrous cells in the gland with the reduction or atrophy of one lung lobe . P. J . DEORAS and N. E. VAD. (Haffkine Institute, Bombay-12) . Vacuum method of milking
and significance of venom out-put data.
Extraction of venom by a vacuum method was found helpful in the case of Bungarus caeruleus. The venom out-put from the same species of snake from different geographical regions was studied for toxicity . Snake bite data for different locations on the body and at different seasons were scrutinized. This was correlated with the differential output in venom in different months . The utility and significance of such data are discussed. G. D. DumTtzov . (Department of Organic Chemistry, Institute of Chemical Technology, Sofia 56, Bulgaria). Isolation and characterization of a non-heme iron-containing glycopeptide from the venom of Vipera russelli. A glyeopeptide containing non-heme iron has been isolated from the venom of the snake Vipera russelli. The purification procedure includes four steps: starch block electrophoresis, gel filtration on Sephadex G-75, ion exchange chromatography on QÀE-Sephadex and dialysis through Visking Co . cellophane tubings, 8/32 . Homogeneity was verified by acrylamide disc electrophoresis and N-terminal amino acid determination . The N-terminal group was found to be alanine. Amino acid analysis and spectral characterization were performed. In the carbohydrate portion after acidic hydrolysis were identified glucose, glucosamine and silaic acid . It was demonstrated by gel filtration under different conditions that this component formed complexes with some high molecular proteins of the venom, being in a state of associationdissociation equilibrium . The glycopeptide revealed a biological activity when tested on mice . This is the first time that such a substance was found in snake venom. M. Dzouic and I. L. BoNTA. (Department of Pharmacology, Medical Faculty Rotterdam, Rotterdam, The Netherlands) . Effect of Naja naja venom and a heparin-precipitable portion on threshold to nocioceptive and convulsive stimuli on rats . Observations concerning the analgesic and anti-epileptic properties of cobra venom were reported in the past but contradictory data in man and laboratory animals were also published (Lusn?vnv and WAD, 1954 ; Wnjt"m, 1960 ; VoRoBEv et al., 1968 ; D'AmouR and SMuTx, 1941). A reappraisal of the question on animal experiments was the aim of the present work. We used crude Naja ngla venom and two portions of it obtained by addition of heparin to the venom solution, and subsequent centrifugation. The two materials thus obtained were the supernatant containing the less basic and the precipitate containing the more basic components of the venom. Heparin was removed by addition of protamine sulphate and subsequent centrifugation . Rats were used throughout and the venom or its portions were administered s .c . As judged by the LD . the supernatant, containing the neurotoxin, was slightly more toxic (0295 mg/kg) and the precipitate considerably less toxic (3-10 mg/kg) than the full venom TOXICON 1972 VoL 10.