Chemical modification of tryptophan residues in alpha-neurotoxins from Ophiophagus hannah (king cobra) venom.
Two alpha-neurotoxins, Oh-4 and Oh-7, from the king cobra (Ophiophagus hannah) venom were subjected to Trp modification with 2-nitrophenylsulfenyl chloride (NPS-Cl). One major NPS derivative was isolated from the modified mixtures of Oh-4 and two from Oh-7 by HPLC. Amino acid analysis and sequence determination revealed that Trp-27 in Oh-4, and Trp-30 and Trp-26 and 30 in the two Oh-7 derivatives, were modified, respectively. Sulfenylation of Trp-27 in Oh-4 caused about 70% drop in lethal toxicity and nicotinic acetylcholine receptor-binding activity. Modification of Trp-30 in Oh-7 resulted in the decrease of lethal toxicity by 36% and binding activity by 61%. The activities were further lost when the conserved Trp-26 in Oh-7 was modified. Sulfenylation of the Trp residues did not significantly affect the secondary structure of the toxins as revealed by the CD spectra. These results indicate that the Trp residues in these two long alpha-neurotoxins may be involved in the receptor binding.
Chang
C C
CC
Department of Biochemistry, Kaohsiung Medical College, Taiwan, ROC.
Lin
P M
PM
Chang
L S
LS
Kuo
K W
KW
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D002038:Bungarotoxins / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D002918:Chymotrypsin / Q000378:metabolism; D002942:Circular Dichroism; D003039:Cobra Neurotoxin Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism / Q000633:toxicity; D007928:Lethal Dose 50; D051379:Mice; D008969:Molecular Sequence Data; D009578:Nitrobenzenes; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification; D011978:Receptors, Nicotinic / Q000378:metabolism; D017421:Sequence Analysis; D013050:Spectrometry, Fluorescence; D014364:Tryptophan / Q000737:chemistry*
D002038:Bungarotoxins; D003039:Cobra Neurotoxin Proteins; D009578:Nitrobenzenes; D010455:Peptides; D011978:Receptors, Nicotinic; C025154:alpha-cobratoxin; C002732:2-nitrobenzenesulfenyl chloride; D014364:Tryptophan; D002918:Chymotrypsin