Improved method for the isolation, characterization and examination of neuromuscular and toxic properties of selected polypeptide fractions from the crude venom of the Taiwan cobra Naja naja atra.
An improved chromatographic method was developed to isolate and purify polypeptides and proteins from the crude venom of the Taiwan cobra Naja naja atra. The procedure devised is simple, easy to reproduce, and enables large scale isolation of almost all polypeptides and proteins in this cobra venom. Six pure polypeptide fractions of the venom were isolated and characterized using gel filtration on Sephadex G50 (medium), ion exchange chromatography on SP-Sephadex C25, desalting on Sephadex G25 (fine) and preparative HPLC on a RPC 18 column. The neuromuscular activity of these fractions was tested on the chick biventer cervicis nerve-muscle preparation and their toxicity (LD(50)) was determined after i.v. administration in mice. Their antinociceptive activity was tested in the mouse abdominal test by i.v. application. Two of these polypeptide samples had major physiological effects: one acted as a cardiotoxin causing reversible myocardial contractures with no effect on muscle twitches elicited by nerve stimulation (NS); another was a neurotoxin that blocked muscle contractions in response to NS and exogenously added acetylcholine. The cardiotoxic fraction was identified as CTX I, a well-known cardiotoxin present in this venom, and the neurotoxin was identified as neurotoxin-α with an LD50 in mice of 0.075 mg/kg.
Hannover Medical School, Clinic for Immunology and Rheumatology, Peptide Chemistry Group, Feodor-Lynen Str. 31, 30625 Hannover, Germany. L.standker@gmx.de
L
L
Ständker
A L
AL
Harvey
S
S
Fürst
I
I
Mathes
W G
WG
Forssmann
G
G
Escalona de Motta
L
L
Béress
D000700:Analgesics / Q000302:isolation & purification / Q000494:pharmacology; D000818:Animals; D005591:Chemical Fractionation / Q000295:instrumentation / Q000379:methods; D002645:Chickens; D002845:Chromatography / Q000379:methods*; D004179:Cobra Cardiotoxin Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000633:toxicity; D003039:Cobra Neurotoxin Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000633:toxicity; D004546:Elapid Venoms / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity; D017815:Elapidae / Q000502:physiology; D005260:Female; D006321:Heart / Q000187:drug effects; D007928:Lethal Dose 50; D008297:Male; D051379:Mice; D009119:Muscle Contraction / Q000187:drug effects; D018482:Muscle, Skeletal / Q000187:drug effects; D009200:Myocardial Contraction / Q000187:drug effects; D009469:Neuromuscular Junction / Q000187:drug effects / Q000503:physiopathology; D010146:Pain / Q000139:chemically induced / Q000188:drug therapy; D017288:Pain Threshold / Q000187:drug effects; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification* / Q000633:toxicity; D051381:Rats
D000700:Analgesics; D004179:Cobra Cardiotoxin Proteins; D003039:Cobra Neurotoxin Proteins; D004546:Elapid Venoms; D010455:Peptides