Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10880761 | Toxicon | 2005 | 7 Pages |
Abstract
Venom was milked by gently pressing the base of the opercular and dorsal fin spines. Three fractions were obtained by molecular exclusion high pressure liquid chromatography (HPLC) (Protein Pak⢠125SW, Millipore Corporation) column, but only the last one with 22.7 min retention time (rt) was biological active (TmPP-22.7). This fraction was rechromatographed on reversed phase HPLC chlorobutylsilane columns (C4, Vydac) nine fractions were obtained, but only one (TmC4-47.2) with 47.2 min rt was biologically active. MALD-TOF mass analysis was carried out on two samples of TmC-47.2 and the results were 15,161.36 and 15,154.70 a.m.u., respectively. Raw venom (1040 μg/ml) depolarised frog (Hyla crepitans) muscle irreversibly from â85 (â88, â81) mV (n=20, median and its 95% CI) to â18 (â24, â15) mV (n=24). The biological activity in TmPP-22.7 (38 μg/ml), which depolarised muscle fibres from â79 (â82, â76) mV (n=20) to â63 (â69 â57) mV (n=24). The depolarising fraction was TmC4-47.2 (50 μg/ml) which depolarised muscles from â87 (91, â82) mV (n=33) to â63 (â76 â51) mV (n=53); the depolarising effect at this concentration was completely reversed on washing with normal saline for 2 h. Muscles treated with 1 μM tetrodotoxin (TTX) were depolarised from â80 (â85, â72) mV (n=49) to â44 (â56, â31) mV (n=44) when 100 μg/ml TmC4-47.2 were applied with TTX; washing 130 min with 1 μM TTX repolarised to â59 (â69, â50) mV (n=25). We also present evidence that TmC4-47.2 induces myonecrosis in mice.
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Authors
J.I. Sosa-Rosales, G. D'Suze, V. Salazar, J. Fox, C. Sevcik,