Data set for the proteomic inventory and quantitative analysis of chicken eggshell matrix proteins during the primary events of eggshell mineralization and the active growth phase of calcification

This research was funded by the French National Research Agency ANR (ANR-13-BSV6-0007-01, ANK-13-BSV6-0007-02 and ANK-13-BSV6-0007-05). The high resolution mass spectrometer was financed (SMHART project, 35069) by the European Regional Development Fund (ERDF), the Conseil Regional du Centre, the French National Institute for Agricultural Research (INRA) and the French National Institute of Health and Medical Research (Inserm), ARN acknowledges funding through Grants CGL2011-25906 (Ministerio de Economia, Spain).Chicken eggshell is a biomineral composed of 95% calcite calcium carbonate mineral and of 3.5% organic matrix proteins. The assembly of mineral and its structural organization is controlled by its organic matrix. In a recent study [1], we have used quantitative proteomic, bioinformatic and functional analyses to explore the distribution of 216 eggshell matrix proteins at four key stages of shell mineralization defined as: (1) widespread deposition of amorphous calcium carbonate (ACC), (2) ACC transformation into crystalline calcite aggregates, (3) formation of larger calcite crystal units and (4) rapid growth of calcite as columnar structure with preferential crystal orientation. The current article detailed the quantitative analysis performed at the four stages of shell mineralization to determine the proteins which are the most abundant. Additionally, we reported the enriched GO terms and described the presence of 35 antimicrobial proteins equally distributed at all stages to keep the egg free of bacteria and of 81 proteins, the function of which could not be ascribed.French National Research Agency (ANR) ANR-13-BSV6-0007-01 ANK-13-BSV6-0007-02 ANK-13-BSV6-0007-05European Union (EU) 35069Region Centre-Val de LoireFrench National Institute for Agricultural Research (INRA)Institut National de la Sante et de la Recherche Medicale (Inserm)Ministerio de Economia, Spain CGL2011-2590

Purification of native HBHA from Mycobacterium avium subsp. paratuberculosis.

International audienceBACKGROUND: Paratuberculosis remains today a major global problem in animal health, especially for dairy cattle. However, the diagnosis of its etiologic agent, Mycobacterium avium subsp. paratuberculosis (Map), still lacks sensitivity because of the lack of available antigens. Little is known about the virulence factors for this pathogen. In this study we have developed a method to produce and purify the heparin-binding hemagglutinin (HBHA), a major adhesin of Mycobacteria, from a culture of Map. FINDINGS: For this extremely slow-growing Mycobacterium, a culture was established in a 3-liter bioreactor. Using the bioreactor the amount of the Map biomass was increased 5-fold compared to a classical culture in flasks. The map-HBHA was purified from a Map lysate by heparin-Sepharose chromatography on HiTrap columns. Binding of map-HBHA onto heparin-Sepharose can be reduced in the presence of salt. Consequently, all steps of sample preparation and column equilibration were carried out in 20 mM Tris-HCl (pH 7.2). The map-HBHA was eluted by a linear NaCl gradient. High resolution mass spectrometry analyses revealed that the native form of map-HBHA has posttranslational modifications, including the removal of the initiation methionine, acetylation of the alanine residue at the N-terminal extremity and the presence of methylated lysines in the C-terminal domain of the protein. CONCLUSIONS: An optimized culture of Map in a bioreactor was established to purify the native map-HBHA from a Map lysate by heparin-Sepharose chromatography. The availability of this antigen offers the possibility to study the structure of the protein and to examine its role in pathogenicity, in particular to better understand the specific interactions of Map with the intestinal tissue. The map-HBHA obtained in its native immunogenic form may also be useful to improve the diagnostic test, especially for the development of a new T-cell-based interferon gamma release assays

B4GALNT2, a new major fecundity gene in the Lacaune ovine breed

National audienc

Qualitative proteomic analysis of Tipula oleracea nudivirus occlusion bodies

Nudiviruses are arthropod-specific large double-stranded circular DNA viruses, related to baculoviruses, which replicate in the nucleus of the cells they infect. Up to date six fully sequenced nudiviral genomes are available in databases and protein profile from nudivirus particles were mainly characterized by polyacrylamide gel electrophoresis. However, only few direct matches were completed between genomic and proteomic data to the exception of the major occlusion body protein from Penaeus monodon nudivirus (PmNV) and four nucleocapsid proteins from Helicoverpa zea nudivirus 2 (HzNV-2). Function of predicted nudiviral proteins is still inferred from what is known from baculoviruses or endogenous nudiviruses (i.e. bracoviruses). Tipula oleracea nudivirus (ToNV) is the causative agent of crane fly nucleopolyhedrosis. With PmNV, ToNV is the second fully sequenced nudivirus to be described as forming occlusion bodies. Protein profile revealed by Coomassie-stained SDS-PAGE is quite similar to those observed for other nudiviruses with five major protein bands of about 75, 48, 35, 25 and 12 kDa. Proteomic analysis using on-line nanoflow liquid chromatography tandem high resolution mass spectrometry revealed ToNV occlusion bodies are composed of 52 viral proteins, the most abundant of which are the functional homolog of baculovirus polyhedrin/granulin and the homologs of three HzNV-2 predicted proteins: the two virion structural proteins 34K (Hz2V052, the baculovirus capsid protein VP39 homolog) and 11K (Hz2V025); and the hypothetical protein Hz2V079, a newly identified nudivirus core gene product

Differential and quantitative analysis of dog oviductal fluid

Poster P69absen

Differential proteomic analysis of bovine oocytes and cumulus cells: zoom on histones abundance and modifications during maturation

International audienc

From single cell MALDI-TOF profiling to protein identification in bovine oocyte: integrative approach to study oocyte maturation

National audienc

Utilization of label free spectral counting to quantify the proteins involved in the various phases of chicken eggshell mineralisation

National audienc

Quantification et caractérisation des protéines sécrétées par l'utérus lors de l'initiation, de la croissance et de l'arrêt de la minéralisation de la coquille d'oeuf de poule

National audienc

Proteomic analysis of the occluded Tipula oleracea nudivirus (ToNV)

International audienc