Purification from black widow spider venom of a protein factor causing the depletion of synaptic vesicles at neuromuscular junctions

The aqueous extract of the venom glands of black widow spiders was fractionated on a column of Sephadex G-200 and then on a column of DEAE-Sephadex A-50, pH 8.2. A protein fraction was obtained that caused a great increase in the frequency of occurrence of miniature end plate potentials at the frog...

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Bibliographic Details
Published in:The Journal of cell biology 1976-03, Vol.68 (3), p.462-479
Main Authors: Frontali, N, Ceccarelli, B, Gorio, A, Mauro, P, Siekevitz, P, Tzeng, M.C, Hurlbut, W.P
Format: Article
Language:English
Subjects:
Animals
Anura
arthropods
Astacoidea - drug effects
Black Widow Spider
Cockroaches - drug effects
Electrophoresis
entomology
Fractionation
Gels
Heart - drug effects
insects
Isoelectric Point
Mechanoreceptors - drug effects
Membrane Potentials - drug effects
Mice
Molecular Weight
Nerves
Neuromuscular Junction - drug effects
Proteins - analysis
Proteins - isolation & purification
Proteins - pharmacology
Proteins - toxicity
Spider venoms
Spiders
Swelling
Synapses
Synaptic Vesicles - drug effects
Toxicity
Venoms
Venoms - analysis
Venoms - pharmacology
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Summary:The aqueous extract of the venom glands of black widow spiders was fractionated on a column of Sephadex G-200 and then on a column of DEAE-Sephadex A-50, pH 8.2. A protein fraction was obtained that caused a great increase in the frequency of occurrence of miniature end plate potentials at the frog neuromuscular junction, and caused swelling of the nerve terminals and depleted them of their vesicles. The fraction consists of at least four protein components that are similar in their molecular weights (about 130,000) and isoelectric points (ranging from pH 5.2 to 5.5) and are immunologically indistinguishable. It contains no sugar residues and has little or no lipolytic or proteolytic activity. The fraction is toxic to mice and is different from the fractions that act on houseflies, the crayfish stretch receptor and the cockroach heart. It seems pure enough to warrant a detailed study of its site and mode of action.
ISSN:0021-9525
1540-8140
DOI:10.1083/jcb.68.3.462