Measurement of vitellogenin protein in invertebrates: relevance and usefulness of mass spectrometry (LC-MS/MS) to propose a specific and transferable method across species - Archive ouverte HAL Accéder directement au contenu
Communication Dans Un Congrès Année : 2011

Measurement of vitellogenin protein in invertebrates: relevance and usefulness of mass spectrometry (LC-MS/MS) to propose a specific and transferable method across species

Résumé

Reproductive success of organisms is related to the quantity and quality of eggs produced by females. Vitellogenin (Vg) is the precursor protein of vitellin that is the energy available for embryonic development in oviparous organisms. Vg is proposed as a relevant biomarker of endocrine disruptors in aquatic vertebrate species (e.g. fish, amphibian). Numerous strategies, such as enzyme-linked immunosorbent assays have been developed to characterize and quantify this protein in vertebrates. On the contrary, in invertebrates few methods are available in spite of their importance. This gap mainly comes from the low transferability of available antibodies across species. Recently, our laboratory developed a quantitative assay of Vg in a widespread amphipod, Gammarus fossarum, using liquid chromatography tandem mass spectrometry (LC-MS/MS) in selected reaction monitoring mode (SRM). In this context, the aim of this study was to assess the possibility to take advantage of peptidic motifs conservation to propose a transferable method across species. For this, our study focused on three model species for which sequence of egg yolk protein is well known: an amphipod (Gammarus fossarum), a cladocerean (Daphnia magna) and an insect (Drosophila melanogaster). In a first step, proteotypic peptides of Vg were identified for each species. Then we tried to find these proteotypic peptides in closely related species for which Vg sequence is unknown (such as G. pulex, G. wautierii, D. pulex). This study showed the high relevance of mass spectrometry to propose a specific methodology for Vg absolute quantification in organisms for which proteins sequences are unknown. Moreover, the proposed methodology is transferable from a species to another one.
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Dates et versions

hal-00960729 , version 1 (18-03-2014)

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  • HAL Id : hal-00960729 , version 1

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Romain Simon, Fabien Audouard-Combe, Guillaume Jubeaux, Renaud Tutundjian, Hervé Quéau, et al.. Measurement of vitellogenin protein in invertebrates: relevance and usefulness of mass spectrometry (LC-MS/MS) to propose a specific and transferable method across species. Congrès français de Spectrométrie de Masse et d'Analyse Protéomique (SMAP 2011), Sep 2011, Avignon, France. ⟨hal-00960729⟩
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