Probing the structural diversities of long alpha-neurotoxins by fluorescence quenching studies.
Trp fluorescence of Ophiophagus hannah neurotoxins (Oh-4, Oh-6A, Oh-7, and Oh-8) and Bungarus multicinctus (alpha-bungarotoxin was quenched by acrylamide and iodide. Acrylamide quenching studies indicated that the degree of exposure of Trp residues in the neurotoxins followed the order Oh-8 > Oh-7 > Oh-6A > Oh-4 > alpha-bungarotoxin, as did the accessibility for iodide. These results reveal that the exposed degree of Trp residues and the microenvironment surrounding Trp residues in the neurotoxins differ, even though their Trp residues and positively charged residues are located at the same or homologous positions. In contrast to unfolded Oh-4, Oh-6A, Oh-7, and alpha-bungarotoxin, unfolding of Oh-8 by reduction and S-carboxymethylation caused a notable decrease in the susceptibility of their Trp residues for iodide. These observations support the view that the side chains of Trp residues and positively charged residues in their native structure do not point toward the same spatial positions. Computer models of the neurotoxins are in good agreement with this proposition. These results elucidate why the conserved Trp residues and cationic groups do not always play the same roles in the biological activities of the neurotoxins.
Chang
L S
LS
Institute of Biomedical Sciences, National Sun Yat-Sen University, Kaohsiung, Taiwan. lschang@mail.nsysu.edu.tw
Lin
S R
SR
Wang
J J
JJ
Chang
C C
CC
D020106:Acrylamide / Q000737:chemistry; D000818:Animals; D017838:Bungarus; D017812:Colubridae; D003198:Computer Simulation; D005453:Fluorescence; D007454:Iodides / Q000737:chemistry; D008958:Models, Molecular; D009498:Neurotoxins / Q000737:chemistry* / Q000302:isolation & purification*; D011487:Protein Conformation; D012910:Snake Venoms / Q000032:analysis*; D013329:Structure-Activity Relationship; D014364:Tryptophan / Q000737:chemistry*
D007454:Iodides; D009498:Neurotoxins; D012910:Snake Venoms; D020106:Acrylamide; D014364:Tryptophan