Differential involvement of disulfide bridges on the folding of a scorpion toxin.
Leiurotoxin I is a neurotoxin, blocker of Ca(2+)-activated apamin-sensitive K+ channel, purified from the venom of the scorpion Leiurus quinquestriatus hebraeus. It is a 31-residue polypeptide reticulated by three disulfide bridges, i.e. Cys3-Cys21, Cys8-Cys26 and Cys12-Cys28. To investigate the role of these disulfide bridges in the folding of this toxin, analogs lacking one disulfide bridge were synthesized. The structures of two analogs in which two half-cystines were placed by alpha-aminobutyrate residues to suppress one disulfide bridge, were analyzed by 1H NMR. The NMR studies reveal a three-dimensional structure identical with the native toxin for the analog lacking disulfide bridge Cys3-Cys21 and a loss of organized structure for another analog lacking disulfide bridge Cys12-Cys28. These analogs are, respectively, fully active and weakly active (2% of the residual activity) when tested in vitro for their ability to interact with their receptor channel and in vivo for their neurotoxic activity in mice. This suggest that disulfide bridge Cys12-Cys28 is essential for the folding process. In contrast, the lack of disulfide bridge Cys3-Cys21 does not affect the folding and the maintenance of bioactive conformation of Leiurotoxin I.
Calabro
V
V
Architecture et Fonction des Macromolécules Biologique, Centre de la Recherche Scientifique, UPR 9039, Marseille, France.
Sabatier
J M
JM
Blanc
E
E
Lecomte
C
C
Van Rietschoten
V
V
Darbon
H
H
D000595:Amino Acid Sequence; D000613:Aminobutyrates / Q000737:chemistry; D000818:Animals; D003553:Cystine / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000737:chemistry / Q000633:toxicity; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity; D011487:Protein Conformation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000378:metabolism; D012604:Scorpion Venoms / Q000737:chemistry* / Q000633:toxicity
D000613:Aminobutyrates; D004220:Disulfides; D009498:Neurotoxins; D010455:Peptides; D011956:Receptors, Cell Surface; D012604:Scorpion Venoms; C056574:leiurotoxin I; D003553:Cystine; C012223:alpha-aminobutyric acid