Purification and sequence determination of a new neutral mammalian neurotoxin from the scorpion Buthus martensii Karsch.
A new neutral mammalian neurotoxin, designated BmK M4, with an isoelectric point (pI) of 7.6 and a relatively low toxicity (LD50 = 4.0 +/- 0.25 microgram/g mice, i.v.) was purified from the venom of scorpion Buthus martensii Karsch (BmK). The complete amino acid sequence of the toxin composed of 64 amino acid residues was determined by automated Edman degradation of the N-terminal part of the reduced and S-carboxamidomethylated protein (up to 30 amino acid residues) and its peptide fragments degraded by lysylendopeptidase or Staphylococcus aureus Va protease. The calculated mol. wt based on the amino acid composition was 7001. By comparison with the sequences of other basic BmK mammalian neurotoxins, it was concluded that the weaker toxicity and lower pI value of BmK M4 might be the result of mutations H10E, R18G and K28D. The sequence comparison of BmK M4 with an acidic toxin, BmK M8, showed that the weakest toxicity and acidic property of BmK M8 may be the consequence of mutations K8D, D53A, V55E and V59E. The substitution of 21 Gly in BmK M4 for Tyrin other BmK toxins may also be of importance. In their tertiary structures, these mutated charged residues are mainly distributed in the surface (face B) that is roughly opposite to the "conserved hydrophobic surface" (face A) proposed by Fontecilla-Camps et al. in 1982. Therefore the toxin-receptor interaction may take a multiposition mode.
Luo
M J
MJ
State Key Laboratory of Molecular Biology, Shanghai Institute of Biochemistry, Academia Sinica, P.R. China.
Xiong
Y M
YM
Wang
M
M
Wang
D C
DC
Chi
C W
CW
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D007928:Lethal Dose 50; D051379:Mice; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000737:chemistry / Q000302:isolation & purification*; D010446:Peptide Fragments / Q000737:chemistry; D012604:Scorpion Venoms / Q000737:chemistry / Q000302:isolation & purification*; D012605:Scorpions
D009498:Neurotoxins; D010446:Peptide Fragments; D012604:Scorpion Venoms