Isolation, partial purification and characterization of active polypeptide from the sea anemone Bartholomea annulata.
In the sea anemone Bartholomea annulata, four different types of cnidocysts, basitrichous isorhizas, microbasic p-mastigophores, microbasic amastigophores and spirocysts were identified. In relation to the efficacy of different substances to induce discharge of nematocysts we observe that distilled water induced more than 70% of microbasic p-mastigophores to discharge, whereas spirocysts were discharged in a lesser extent (approximately 20%). The median lethal dose (LD50) in mice was found after injection of 700.7 mg protein per kg of body weight from the crude extract. The protein with neurotoxic effect was isolated using low-pressure liquid chromatography. The neurotoxic activity was determined using sea crabs (Ocypode quadrata), injecting 15 microg of crude extract or isolated fraction into the third walking leg, and violent motor activity followed by progressive loss of sensibility to external stimuli, further leading to full paralysis were observed. The active fraction (called V) eluted at 43.9 min.
Sánchez-Rodríguez
Judith
J
Institute of Marine Sciences and Limnology, Puerto Morelos Academic Unit, National Autonomous University of Mexico, Puerto Morelos, Mexico. judithsa@mar.icmyl.unam.mx
Zugasti
Alejandro
A
Santamaría
Abel
A
Galván-Arzate
Sonia
S
Segura-Puertas
Lourdes
L
D000818:Animals; D003386:Brachyura / Q000187:drug effects; D002850:Chromatography, Gel / Q000379:methods; D003064:Cnidarian Venoms / Q000737:chemistry / Q000302:isolation & purification* / Q000633:toxicity; D005612:Freeze Drying / Q000379:methods; D007274:Injections, Intraperitoneal; D007928:Lethal Dose 50; D008297:Male; D051379:Mice; D009043:Motor Activity / Q000187:drug effects; D009498:Neurotoxins / Q000737:chemistry / Q000302:isolation & purification / Q000633:toxicity; D010243:Paralysis / Q000139:chemically induced / Q000401:mortality; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification* / Q000633:toxicity; D012615:Sea Anemones / Q000737:chemistry / Q000378:metabolism*; D023382:Toxicity Tests, Acute / Q000379:methods; D014202:Tremor / Q000139:chemically induced
D003064:Cnidarian Venoms; D009498:Neurotoxins; D010455:Peptides