title: The N-terminal amino group essential for the biological activity of notexin from Notechis scutatus scutatus venom.
abstract: Notexin from Notechis scutatus scutatus snake venom was modified with trinitrobenzenesulfonic acid, and the major trinitrophenylated (TNP) derivative was separated by high-performance liquid chromatography. Modification resulted in the incorporation of only one TNP group on the N-terminal alpha-amino group. The TNP derivative showed a precipitous decrease in enzymatic activity and lethal toxicity, whereas the antigenicity remained unchanged. However, trinitrophenylation did not significantly affect the secondary structure of the toxin molecule as revealed by the CD spectra. The results, that the modification reaction was accelerated by the Ca2+ and that the TNP derivative retains its affinity for Ca2+, indicate that the N-terminal alpha-amino group did not participate in the Ca2(+)-binding. The TNP derivative could be regenerated with hydrazine hydrochloride. The biological activities of the regenerated notexin are almost the same as those of native notexin. These results suggest that the N-terminal alpha-amino group is essential for the phospholipase A2 activity and lethal toxicity of notexin, and that incorporation of the TNP group on the N-terminal alpha-amino group might give rise to a distortion of the active conformation of notexin.
authors.affiliation: Institute of Life Sciences, National Tsing Hua University, Hsinchu, Taiwan, China.
authors.forename: C C
authors.initials: CC
authors.lastname: Yang
authors.forename: L S
authors.initials: LS
authors.lastname: Chang
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002118:Calcium / Q000494:pharmacology; D002852:Chromatography, Ion Exchange; D002942:Circular Dichroism; D004546:Elapid Venoms / Q000302:isolation & purification / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009498:Neurotoxins; D011487:Protein Conformation; D014302:Trinitrobenzenesulfonic Acid
chemical_list: D004546:Elapid Venoms; D009498:Neurotoxins; C005082:notexin; D014302:Trinitrobenzenesulfonic Acid; D002118:Calcium