Determination of the disulfide bond pattern of a novel C-type lectin from snake venom by mass spectrometry.
The disulfide bond pattern of Trimeresurus stejnegeri lectin (TSL), a new member of the C-type lectin family, was determined by mass spectrometry. Four intrachain disulfide bonds of TSL, Cys(3)-Cys(14), Cys(31)-Cys(131), Cys(38)-Cys(133) and Cys(106)-Cys(123), and two interchain linkages, Cys(2)-Cys(2) and Cys(86)-Cys(86), were determined. Three strategies were used in this work. One intrachain (Cys(106)-Cys(123)) and one interchain (Cys(86)-Cys(86)) disulfide linkages were detected by standard MS methods. The disulfide bonds Cys(2)-Cys(2) and Cys(3)-Cys(14) were analyzed using a modified partial reduction procedure and MS/MS. The last two disulfide bonds were characterized by a MS/MS/MS technique. The strategies developed in this work could be applied more generally to detection of disulfide bond patterns.
Zeng
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Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, People's Republic of China.
Xu
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Shao
X X
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Wang
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Xia
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QC
D000595:Amino Acid Sequence; D002853:Chromatography, Liquid; D003435:Crotalid Venoms / Q000032:analysis / Q000737:chemistry*; D003545:Cysteine / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry*; D037102:Lectins / Q000032:analysis / Q000737:chemistry*; D037181:Lectins, C-Type; D008969:Molecular Sequence Data; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D003435:Crotalid Venoms; D004220:Disulfides; D037102:Lectins; D037181:Lectins, C-Type; C029748:Trimeresurus venoms; D003545:Cysteine