One-disulfide intermediates of apamin exhibit native-like structure.
The conformations of three peptide models of the one-disulfide and fully reduced forms of apamin were characterized by using nuclear magnetic resonance (NMR) spectroscopy. Apa-2 contains the native disulfide bond between Cys3 and Cys15 in apamin with the other two cysteines replaced by alanines. Apa-1 contains the native disulfide bond between Cys1 and Cys11. Apa-S has all cysteines replaced with serines, mimicking fully reduced apamin. Comparing NOESY cross peaks and coupling constants for amide protons in the peptide analogs with those in native apamin indicates that a significant portion of Apa-2 possesses native-like structural elements of apamin in addition to some random coil conformations. Apa-1 contains a short helical structure from Ala9 to Arg13, corresponding to the N-terminal portion of the alpha-helix observed in the native structure, along with some local and probably flexible secondary structures corresponding to the reverse turn region in native apamin. A larger portion of Apa-1 exists in the form of random coil conformations compared to Apa-2. Apa-S displays mainly random coil conformations with some localized helical structures from Glu7 to Arg14 which are similar to the "nascent helices" proposed by Wright et al. [Wright, P. E., Dyson, H. J., & Lerner, R. A. (1988) Biochemistry 27, 7167-7175]. Formation of the first disulfide bond in apamin seems to be important in the folding process by stabilizing native-like structure, presumably by reducing the conformational freedom and initiating formation of structure.(ABSTRACT TRUNCATED AT 250 WORDS)
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Department of Biochemistry, Louisiana State University, Baton Rouge 70803.
Nelson
J W
JW
D000595:Amino Acid Sequence; D001030:Apamin / Q000031:analogs & derivatives / Q000737:chemistry*; D004220:Disulfides; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D010456:Peptides, Cyclic / Q000138:chemical synthesis / Q000737:chemistry*; D011487:Protein Conformation; D017433:Protein Structure, Secondary
D004220:Disulfides; D010455:Peptides; D010456:Peptides, Cyclic; D001030:Apamin