| Title |
Assigning Peptide Disulfide Linkage Pattern Among Regio-Isomers via Methoxy Addition to Disulfide and Tandem Mass Spectrometry
|
|---|---|
| Published in |
Journal of the American Society for Mass Spectrometry, February 2017
|
| DOI | 10.1007/s13361-017-1595-1 |
| Pubmed ID | |
| Authors |
Kirt L. Durand, Lei Tan, Craig A. Stinson, Chasity B. Love-Nkansah, Xiaoxiao Ma, Yu Xia |
| Abstract |
Pinpointing disulfide linkage pattern is critical in the characterization of proteins and peptides consisting of multiple disulfide bonds. Herein, we report a method based on coupling online disulfide modification and tandem mass spectrometry (MS/MS) to distinguish peptide disulfide regio-isomers. Such a method relies on a new disulfide bond cleavage reaction in solution, involving methanol as a reactant and 254 nm ultraviolet (UV) irradiation. This reaction leads to selective cleavage of a disulfide bond and formation of sulfenic methyl ester (-SOCH3) at one cysteine residue and a thiol (-SH) at the other. Under low energy collision-induced dissociation (CID), cysteine sulfenic methyl ester motif produces a signature methanol loss (-32 Da), allowing its identification from other possible isomeric structures such as S-hydroxylmethyl (-SCH2OH) and methyl sulfoxide (-S(O)-CH3). Since disulfide bond can be selectively cleaved and modified upon methoxy addition, subsequent MS(2) CID of the methoxy addition product provides enhanced sequence coverage as demonstrated by the analysis of bovine insulin. More importantly, this reaction does not induce disulfide scrambling, likely due to the fact that radical intermediates are not involved in the process. An approach based on methoxy addition followed by MS(3) CID has been developed for assigning disulfide linkage patterns in peptide disulfide regio-isomers. This methodology was successfully applied to characterizing peptide systems having two disulfide bonds and three disulfide linkage isomers: side-by-side, overlapped, and looped-within-a-loop configurations. Graphical Abstract ᅟ. |
X Demographics
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Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Scientists | 1 | 100% |
Mendeley readers
The data shown below were compiled from readership statistics for 16 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 16 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 3 | 19% |
| Researcher | 3 | 19% |
| Student > Doctoral Student | 2 | 13% |
| Professor | 2 | 13% |
| Student > Bachelor | 2 | 13% |
| Other | 2 | 13% |
| Unknown | 2 | 13% |
| Readers by discipline | Count | As % |
|---|---|---|
| Chemistry | 6 | 38% |
| Biochemistry, Genetics and Molecular Biology | 1 | 6% |
| Mathematics | 1 | 6% |
| Nursing and Health Professions | 1 | 6% |
| Pharmacology, Toxicology and Pharmaceutical Science | 1 | 6% |
| Other | 4 | 25% |
| Unknown | 2 | 13% |
Attention Score in Context
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 04 March 2017.
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#22,764,772
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Outputs from Journal of the American Society for Mass Spectrometry
#3,431
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#371,684
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Outputs of similar age from Journal of the American Society for Mass Spectrometry
#51
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