La présence aux E.D est obligatoire. On the other hand, MEF KO cells expressing the S370A mutant had an enhanced proliferation rate when compared with WT Grb14 (53%, P < 0.01), although it remained much lower than in control KO cells (P < 0.001). 2011 - 2014. Faculté de médecine Paris Descartes ... (DFGSM2 et DFGSM3) ne sont pas autorisés à s’inscrire au PIR MIcrobiologie. Organisation de l’enseignement They also contain a so-called BPS (between PH and SH2 domains) region, also known as PIR (phosphorylated insulin receptor interacting region) that is unique to this family of proteins. ... M1/PIR en Hématologie, étudiante en 4ème année de pharmacie. This work was supported by grants from the Agence Nationale de la Recherche (ANR-06-PHYSIO-014-01), the Association pour la Recherche sur le Cancer (ARC 3487) and “La Ligue Contre Le Cancer,” Comité du Nord, 2006, 2008. Immunoreactive bands were revealed using the ECL detection kit (Pierce, Rockford, IL). Detection of β2-adrenergic receptor dimerization in living cells using bioluminescence resonance energy transfer (BRET). As mentioned above, R387 is also in close proximity to the K1168 IR residue. Lors de sa 5e année, l’année hospitalo-universitaire est maintenue en limitant le choix des stages à des laboratoires hospitaliers dont la vocation est la recherche orientée clinique (centre d’investigation clinique, centre d’investigation biomédical...). As measured in the MEF KO proliferation assay, the R385N mutation strikingly decreased the inhibitory effect of Grb14, because the increase in cell number in the proliferation assay was 3.2-fold higher in KO cells transfected with this mutant compared with WT Grb14. These data thus give clear evidence that the IR αC-helix is involved in Grb14 binding. All chemicals were from Sigma-Aldrich (St. Louis, MO). bioluminescence resonance energy transfer; phosphorylated insulin receptor interacting region; Asante-Appiah E, Kennedy BP2003 Protein tyrosine phosphatases: the quest for negative regulators of insulin action. Camille Taccola. Si vous rencontrez un problème avec le lien du portail, agrandissez votre fenêtre. — Les candidatures pour l’année universitaire 2019-2020 sont ouvertes en ligne du 20 avril jusqu’au 31 mai 2020. Alterations in insulin signaling and action lead to pathophysiological states such as obesity and type 2 diabetes, which are worldwide expanding diseases. We first verified Grb14’s ability to bind to PDK1 and the implication of the E193 residue in this interaction in Xenopus oocytes. Ce stage est suivi d’un mémoire que l’étudiant doit défendre, en partie en anglais, devant un jury. Ils permettent d’approfondir les outils mathématiques et statistiques, ainsi que d’appréhender la chimie biologique. In addition to the interaction of the pseudosubstrate region, the crystallographic study of the BPS/IRK complex identified R387 in Grb14 (R387 in the human sequence, corresponding to R385 in the rat sequence) as a key residue stabilizing the conformation of this region (13). Among the Grb7 family of adapters, Grb14 is likely to be particularly involved in the regulation of insulin signaling, as assessed by its tissue expression (8, 11), by its inhibitory effect on IR catalytic activity (12), by its co-internalization with the IR in insulin-stimulated rat liver (10), and by the phenotype of mice deficient for the Grb14 gene (9). Evidence for a direct interaction between insulin receptor substrate-1 and Shc. 5C). Les étudiants en pharmacie sont sélectionnés sur dossier et entretien dès le début de la 2e année. — Pour pré-candidater en masters, en formation initiale ou formation continue, veuillez utiliser l’application de candidatures en ligne de l’Université Paris Descartes. Effects of mutations of the insulin receptor on its interaction with Grb14. 3D, Grb14 inhibition of insulin-induced oocyte maturation was almost abolished by the nonphosphorylatable S370A mutant. Indeed, Grb14 expression level in white adipose tissue is negatively correlated with insulin sensitivity (8), and Grb14-deficient mice exhibit improved glucose homeostasis and enhanced insulin signaling (9). Paris et … Liste des pays concernés : Algérie, Argentine, Bénin, Brésil, Burkina Faso, Burundi, Cameroun, Chili, Chine, Colombie, Comores, Congo, Corée du Sud, Côte d’Ivoire, Djibouti, Égypte, États-Unis, Gabon, Guinée, Inde, Indonésie, Iran, Japon, Koweït, Liban, Madagascar, Mali, Maroc, Maurice, Mauritanie, Mexique, Pérou, République démocratique du Congo, Russie, Sénégal, Singapour, Taïwan, Togo, Tunisie, Turquie, Vietnam. Among them, it was reported that PDK1 interacted constitutively with the E193 residue of Grb14 and that this interaction was required for a correct activation of Akt in response to insulin (14). Results are means ± sd of three independent animals, each concerning 20 oocytes. Upon autophosphorylation and activation of the IRK, two domains are subjected to major conformational changes: the activation loop of the kinase, which bears the three phosphorylated tyrosine residues Y1158, Y1162, and Y1163, and the αC-helix (amino acids 1038–1054) whose residues become more solvent exposed (17). Thus, PDK1 recruitment is likely to be an important component of the Grb14 inhibitory mechanism. Twenty treated oocytes were lysed in 200 μl buffer [25 mm MOPS (pH adjusted to 7.2), 60 mm β-glycerophosphate, 15 mm paranitrophenyl phosphate, 15 mm EDTA, 15 mm MgCl2, 2 mm dithiothreitol, 1 mm orthosodium vanadate, 1 mm NaF, 1 mm phenylphosphate, 10 μg/ml leupeptin, 10 μg/ml aprotinin, 10 μg/ml soybean trypsin inhibitor, and 10 μm benzamidine] added with 0.5% Triton X-100. This is the first evidence that inhibition by Grb14 of IR tyrosine kinase is not sufficient to inhibit insulin signaling. All together, these data suggest that the R385N mutation drastically reduces the affinity of Grb14 for the IR. Disclosure Summary: The authors have nothing to disclose. We thank Dr. Marthe Rizk-Rabin for her help in proliferation experiments and the technical facilities of the Institut Cochin for flow cytometry analysis and DNA sequencing. Besoin d'aide, contactez-nous via l'adresse . Microinjection of increasing amounts of GST-Grb14 showed that whereas 15 ng had no effect, 20 ng Grb14 totally blocked insulin-induced oocyte maturation.

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