The isolation and purification of two peptides from the venom of Buthus martensii Karsch.
Two peptides that extensively prolong action potentials (APs) in rat and frog nerves have been isolated and purified from the venom of the scorpion Buthus martensii Karsch (BMK). The peptides were purified using gel filtration, ion exchange, FPLC, and HPLC chromatography. Action potentials recorded in the presence of nanomolar concentrations of the peptides were extensively prolonged without much attenuation in their heights. The N-terminal sequences of both the peptides, BMK 9(3)-1 and BMK 9(3)-2, were determined. The N-terminal sequences of BMK 9(3)-1 and BMK 9(3)-2 were found to be: GRDAYIADSEN-PYF-GANPN and GRDAYIADSEN-PYT-ALNP. Sequence similarity comparisons to other alpha-scorpion toxins suggest that the two blanks in each of the sequences are cysteines. The first 20 residues of the two BMK peptides differ by only three amino acid substitutions. The molecular weight (MW) of BMK 9(3)-1 and BMK 9(3)-2 were determined by LC/MS/MS to be 7020 and 7037 Da. Since both of the peptides prolong APs when both K(+) and Ca(++) channels are blocked and show sequence similarity to other alpha-neurotoxins, it appears likely that BMK 9(3)-1 and BMK 9(3)-2 act to alter Na channel inactivation to produce their effects. The first 20 residues of BMK 9(3)-2 are identical to those observed for makatoxin I, a toxin isolated from Buthus martensii Karsch venom, that alters nitric oxide transmitter release. Since the two toxins also have very similar molecular weights, BMK 9(3)-2 may be identical to makatoxin I; however, BMK 9(3)-2 acts to alter Na channels to exert its effect, thus the two toxins may differ, or if they are identical, they can exert effects on both neural transmission and AP propagation.
Chen
Z
Z
Department of Biological Sciences, Northern Illinois University, DeKalb 60115, USA.
Reddy
G
G
Hahin
R
R
D000200:Action Potentials / Q000187:drug effects; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D051381:Rats; D012584:Sciatic Nerve / Q000187:drug effects / Q000502:physiology; D012604:Scorpion Venoms / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D012605:Scorpions; D017386:Sequence Homology, Amino Acid
D000596:Amino Acids; D010455:Peptides; D012604:Scorpion Venoms