Purification and cDNA cloning of an insecticidal protein from the venom of the scorpion Orthochirus scrobiculosus.
Injection of crude venom from the scorpion Orthochirus scrobiculosus into larvae of Heliothis virescens (Lepidoptera: Noctuidae) caused trembling and uncoordinated movement before development of a progressive and prolonged flaccid paralysis. The isolation of the toxin (OsI-1) responsible for this effect of O. scrobiclosus venom is described. The molecular mass of OsI-1 toxin was 6994 Da, as determined by desorption mass spectroscopy. The complete primary structure of OsI-1 was deduced from the sequence of cDNA clones obtained by rapid amplification of cDNA ends (RACE) PCR. Comparison of the deduced amino acid sequence of OsI-1 with those of other insecticidal scorpion toxins indicates that it is a sodium (Na+) channel active depressant insect-selective toxin. The analysis of amino acid sequence of the toxin in conjunction with mass spectroscopy data indicates post-translational modification in maturation with the removal of 3 C-terminal amino acids and amidation of the C-terminus.
Kozlov
S
S
Laboratory of Neuroreceptors & Neuroregulators, Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow.
Lipkin
A
A
Nosyreva
E
E
Blake
A
A
Windass
J D
JD
Grishin
E
E
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002268:Carboxypeptidases / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D006868:Hydrolysis; D007202:Indicators and Reagents; D007306:Insecticides / Q000737:chemistry / Q000302:isolation & purification* / Q000633:toxicity; D007915:Lepidoptera; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000737:chemistry / Q000302:isolation & purification*; D014176:Protein Biosynthesis; D011506:Proteins / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012604:Scorpion Venoms / Q000737:chemistry* / Q000302:isolation & purification* / Q000633:toxicity; D012605:Scorpions / Q000378:metabolism*; D017421:Sequence Analysis; D012997:Solvents; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin
D007202:Indicators and Reagents; D007306:Insecticides; D009498:Neurotoxins; C404752:OsI-1 toxin; D011506:Proteins; D012604:Scorpion Venoms; D012997:Solvents; D002268:Carboxypeptidases; D014357:Trypsin