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Content archived on 2024-04-30

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

Objective



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.

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RGI - Research grants (individual fellowships)

Coordinator

Universidad de Valladolid
EU contribution
No data
Address
7,C/ Ramon y Cajal
47005 Valladolid
Spain

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Total cost

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Participants (1)

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