Purification and characterization of Heteroscorpine-1 (HS-1) toxin from Heterometrus laoticus scorpion venom.
Crude venom from Thai giant scorpion, Heterometrus laoticus, most commonly found in the northeastern area of Thailand, was evaluated for PD50 (paralytic dose 50) activities from abdominal injection to cricket (Gryllus sp.) and activities against various kinds of microorganisms. It exhibited good results in disc diffusion assay for Bacillus subtilis, Klebsiella pneumoniae, Pseudomonas aeruginosa and Staphylococcus aureus. After purification, toxin showed acceptable results for PD50 determination in entrapped cricket as well as inhibitory activity against B. subtilis, K. pneumoniae, and P. aeruginosa with activities over 300 times higher than that of the crude venom. The purified fraction was showed to contain a single band in SDS-PAGE. MALDI-TOF-MS/MS analysis showed one peak of major protein with 8293Da. Partial amino acid sequence show high homology to Scorpine-a polypeptide toxin family with potassium channel blocking and defensin activity. The novel toxin was named "Heteroscorpine-1" (HS-1) as the first Scorpine from genus Heterometrus. After full length determination by PCR, HS-1 gene was found to be composed of two exons, separated by an intron. Deduction revealed 95 amino acid residues with 19 residues as the leading sequence. It showed about 80% similarity to Panscorpine and Opiscorpine.
Uawonggul
Nunthawun
N
Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand.
Thammasirirak
Sompong
S
Chaveerach
Arunrat
A
Arkaravichien
Tarinee
T
Bunyatratchata
Wandee
W
Ruangjirachuporn
Wipaporn
W
Jearranaiprepame
Pornpimol
P
Nakamura
Takeshi
T
Matsuda
Michiyuki
M
Kobayashi
Michimoto
M
Hattori
Seisuke
S
Daduang
Sakda
S
D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000935:Antifungal Agents / Q000494:pharmacology; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis; D017931:DNA Primers; D006090:Gram-Negative Bacteria / Q000187:drug effects / Q000648:ultrastructure; D006094:Gram-Positive Bacteria / Q000187:drug effects / Q000648:ultrastructure; D007267:Injections; D007313:Insecta / Q000187:drug effects; D007928:Lethal Dose 50; D008826:Microbial Sensitivity Tests; D003904:Mitosporic Fungi / Q000187:drug effects; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012604:Scorpion Venoms / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology / Q000633:toxicity*; D012605:Scorpions; D017386:Sequence Homology, Amino Acid
D000900:Anti-Bacterial Agents; D000935:Antifungal Agents; D017931:DNA Primers; D012604:Scorpion Venoms; C517468:heteroscorpine-1, Heterometrus laoticus; D004247:DNA