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Mapping Proteoforms and Protein Complexes From King Cobra Venom Using Both Denaturing and Native Top-down Proteomics
| Content Provider | Scilit |
|---|---|
| Author | Melani, Rafael D. Skinner, Owen S. Fornelli, Luca Domont, Gilberto B. Compton, Philip D. Kelleher, Neil L. |
| Copyright Year | 2016 |
| Description | Journal: Molecular & Cellular Proteomics Characterizing whole proteins by top-down proteomics avoids a step of inference encountered in the dominant bottom-up methodology when peptides are assembled computationally into proteins for identification. The direct interrogation of whole proteins and protein complexes from the venom of Ophiophagus hannah (king cobra) provides a sharply clarified view of toxin sequence variation, transit peptide cleavage sites and post-translational modifications (PTMs) likely critical for venom lethality. A tube-gel format for electrophoresis (called GELFrEE) and solution isoelectric focusing were used for protein fractionation prior to LC-MS/MS analysis resulting in 131 protein identifications (18 more than bottom-up) and a total of 184 proteoforms characterized from 14 protein toxin families. Operating both GELFrEE and mass spectrometry to preserve non-covalent interactions generated detailed information about two of the largest venom glycoprotein complexes: the homodimeric l-amino acid oxidase (∼130 kDa) and the multichain toxin cobra venom factor (∼147 kDa). The l-amino acid oxidase complex exhibited two clusters of multiproteoform complexes corresponding to the presence of 5 or 6 N-glycans moieties, each consistent with a distribution of N-acetyl hexosamines. Employing top-down proteomics in both native and denaturing modes provides unprecedented characterization of venom proteoforms and their complexes. A precise molecular inventory of venom proteins will propel the study of snake toxin variation and the targeted development of new antivenoms or other biotherapeutics. |
| Related Links | https://www.mcponline.org/content/mcprot/15/7/2423.full.pdf https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4937514/pdf http://www.mcponline.org/article/S153594762033526X/pdf |
| Ending Page | 2434 |
| Page Count | 12 |
| Starting Page | 2423 |
| ISSN | 15359476 |
| DOI | 10.1074/mcp.m115.056523 |
| Journal | Molecular & Cellular Proteomics |
| Issue Number | 7 |
| Volume Number | 15 |
| Language | English |
| Publisher | Elsevier BV |
| Publisher Date | 2016-07-01 |
| Access Restriction | Open |
| Subject Keyword | Journal: Molecular & Cellular Proteomics Biochemical Research Mass Spectrometry Protein Identification* Protein Complex Analysis Native Mass Spectrometry Protein Complexes Top-down Proteomics |
| Content Type | Text |
| Resource Type | Article |
| Subject | Medicine Analytical Chemistry Molecular Biology Biochemistry |
