期刊
JOURNAL OF PROTEOME RESEARCH
卷 12, 期 9, 页码 4122-4135出版社
AMER CHEMICAL SOC
DOI: 10.1021/pr400444m
关键词
de novo sequencing; selected reaction monitoring (SRM); targeted proteomics; vitellogenin; isoform identification; egg yolk proteins; sea turtle reproduction
资金
- Agence Nationale de la Recherche [ANR-07-JCJC-0122]
- Fondation pour la Recherche Medicale (FRM) [DGE20101221273]
- Centre National de la Recherche Scientifique (CNRS)
- Bruker Daltonics Company
- French Ministry of Research
- French Proteomics Infrastructure (ProFi) [ANR-10-INSB-08-03]
- Agence Nationale de la Recherche (ANR) [ANR-07-JCJC-0122] Funding Source: Agence Nationale de la Recherche (ANR)
No biomarker has yet been discovered to identify the reproductive status of the endangered leatherback sea turtle (Dermochelys coriacea). Although vitellogenin (VTG) could be used for this, its sequence is not known in D. coriacea and no quantitative assay has been carried out in this species to date. Using de novo sequencing-based proteomics, we unambiguously characterized sequences of two different VTG isoforms that we named Dc-VTG1 and Dc-VTG2. To our knowledge, this is the first clear evidence of different VTG isoforms and the structural characterization of derived yolk proteins in reptiles. This work illustrates how massive de novo sequencing can characterize novel sequences when working on exotic nonmodel species in which even nucleotide sequences are not available. We developed assays for absolute quantitation of these two isoforms using selected reaction monitoring (SRM) mass spectrometry, thus providing the first SRM assays developed specifically for a nonsequenced species. Plasma levels of Dc-VTG1 and Dc-VTG2 decreased as the nesting season proceeded, and were closely related to the increased levels of reproductive effort. The SRM assays developed here therefore provide an original and efficient approach for the reliable monitoring of reproduction cycles not only in D. coriacea, but potentially in other turtle species.
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