A quick procedure for the isolation of dimeric piratoxins-I and II, two myotoxins from Bothrops pirajai snake venom. N-terminal sequencing.
Two myotoxins, MP-I and MP-II, from Bothrops pirajai snake venom, have been purified by a quick high performance liquid chromatography (HPLC) procedure. Based on the HPLC coelution profile, amino acid composition, N-terminal sequence, polyacrylamide gel electrophoresis (PAGE) migration, as well as lack of phospholipase-A2 (PLA2) and proteolytic activities, MP-I and MP-II were identified as piratoxin-I (PrTX-I) and II (PrTX-II), respectively. This procedure affords, aside the reduced operation time, a high yield (35% of the applied sample in terms of A280nm) of MP-I, which is the major myotoxin of the venom. The N-terminal sequences of MP-I, MP-II, PrTX-I and PrTX-II, up to the 51st, 41st, 46th and 39th residues, respectively, have been determined, revealing MP-I (and hence PrTX-I) as a Lys-49 PLA2-like myotoxin. Both MP-I and MP-II have been shown, by SDS-PAGE, to occur in dimeric isoforms.
Toyama
M H
MH
Departamento de Bioquímica, Unicamp, Instituto de Biologia, Brasil.
Mancuso
L C
LC
Giglio
J R
JR
Novello
J C
JC
Oliveira
B
B
Marangoni
S
S
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D054501:Group II Phospholipases A2; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000737:chemistry / Q000302:isolation & purification*; D010741:Phospholipases A / Q000737:chemistry / Q000302:isolation & purification*; D054467:Phospholipases A2; D030162:Reptilian Proteins; D013997:Time Factors
D000596:Amino Acids; D009498:Neurotoxins; D030162:Reptilian Proteins; D010741:Phospholipases A; D054501:Group II Phospholipases A2; D054467:Phospholipases A2; C076780:myotoxin I; C061056:myotoxin II, Bothrops asper