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Contenu archivé le 2024-04-30

Monitoring calcium concentration in the endoplasmic reticulum using recombinant aequorin specifically targeted

Objectif



The project proposed constitutes an extension of that carried out during my stay in the University of Padova. I will first construct a set up to measure aequorin luminescence, similar to those actually running in Padova. Then, I will use the HeLa clones I have obtained in Padova, which stably express the DNA construct designed to target aequorin (both native and mutated with low Ca2+ affinity) to the endoplasmic reticulum (ER). These cells will be used to continue the study of ER-Ca2+-heterogeneity currently being carried out in Padova. At the same time, I plan to produce a series of point mutations in the Ca2+ -binding sites of aequorin in order to reduce its Ca2+-affinity up to near the mM range. In the mean time, Sr2+ will be used as a low-affinity Ca2+-surrogate. Sr2+ has been shown to behave similarly to Ca2+ in terms of uptake, accumulation and release from the ER.

Champ scientifique (EuroSciVoc)

Programme(s)

Thème(s)

Appel à propositions

Données non disponibles

Régime de financement

RGI - Research grants (individual fellowships)

Coordinateur

Universidad de Valladolid
Contribution de l’UE
Aucune donnée
Adresse
7,C/ Ramon y Cajal
47005 Valladolid
Espagne
Coût total
Aucune donnée

Participants (1)