AI Article Synopsis
- The study focused on identifying disulfide bonds in the galactose-specific lectin SEL 24K from Chinook salmon using mass spectrometry techniques.
- Four in silico tools were utilized to predict the oxidation states of cysteine residues and potential locations of the disulfide bonds.
- The researchers confirmed the locations of seven disulfide bonds and a pair of free cysteines through various methods, including digestion, HPLC, mass spectrometry, and fragmentation studies, finding some discrepancies with predictions from public algorithms.
Article Abstract
The disulfide bonds in the galactose-specific lectin SEL 24K from the egg of the Chinook salmon Oncorhynchus tshawytscha were determined by mass spectrometry. Four predictive in silico tools were used to determine the oxidation state of cysteines in the sequence and possible location of the disulfide bonds. A combination of tryptic digestion, HPLC separation, and chemical modifications were used to establish the location of seven disulfide bonds and one pair of free cysteines. After proteolysis, peptides containing one or two disulfide bonds were identified by reduction and mass spectral comparison. MALDI mass spectrometry was supported by chemical modification (iodoacetamide) and in silico digestion. The assignments of disulfide bonds were further confirmed by mass spectral fragmentation studies including in-source dissociation (ISD) and collision-induced dissociation (CID). The experimentally determined disulfide bonds and free Cys residues were only partially consistent with those generated by several automated public-domain algorithms.
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| http://dx.doi.org/10.1016/j.abb.2007.02.002 | DOI Listing |
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