Determination of disulfide bridge pattern in omega-conopeptides.
Synthetic versions of seven naturally occurring omega-conopeptides were subjected to structural analyses in order to determine their disulfide bridge pattern. The method applied in this study uses a combination of amino-acid composition and peptide sequence analysis of various peptide fragments generated by different enzymatic digestions. A temperature modification in the Edman degradation cycles of a protein sequencer allowed the unambiguous detection of the cleavage of cystine residues. The appearance of the cystine residues in particular cycles of the sequence analysis was characteristic of one or several of the theoretically possible 15 isomers. In the case of multiple choices, possible isomers were further eliminated by the amino-acid and sequence analysis of peptide fragments generated by the enzymatic digestion. All synthetic peptides, SNX-111, -157, -159, -183, -185, -230 and -231, were found to have the same disulfide bridge pattern as determined for the naturally occurring omega-conopeptide G-VI-A, i.e. disulfide bridges between the half-cystines 1-16, 8-20 and 15-25 (using the amino-acid numbering of SNX-111).
Chung
D
D
Neurex Corporation, Menlo Park, California, USA.
Gaur
S
S
Bell
J R
JR
Ramachandran
J
J
Nadasdi
L
L
D000595:Amino Acid Sequence; D002121:Calcium Channel Blockers / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D003553:Cystine / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry*; D010450:Endopeptidases / Q000378:metabolism; D008969:Molecular Sequence Data; D008978:Mollusk Venoms / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D020960:omega-Conotoxins
D002121:Calcium Channel Blockers; D004220:Disulfides; D008978:Mollusk Venoms; D010446:Peptide Fragments; D010455:Peptides; D020960:omega-Conotoxins; C087968:omega-conotoxin TVIA; C077380:omega-conotoxin SVIB; C075851:omega-conotoxin-MVIIC; D003553:Cystine; C078452:ziconotide; D010450:Endopeptidases