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Structural basis for Ca2+-transport by the sarcoplasmic Ca2+-pump protein

Objective



The proposal aims at a concerted effort to explore the molecular mechanism of sarcoplasmic reticulum Ca2+-ATPase, a representative of the P type cation transporting ATPases. Ca2+-ATPase structure will be studied by the use of proteolytic dissection, immunolocalization, and chemical modification experiments to define the location of membrane buried and exposed segments of the polypeptide chain and Ca2+ transport sites. Molecular biology tools will be used to develop a yeast system to express relatively large amounts of Ca2+-ATPase mutants to pinpoint amino acid residues involved in Ca2+-binding. Functionally oriented studies will include examination of Ca2+-binding and -release by rapid kinetic experiments. In addition domain organization and proximity-relationships of strategic sites (ATP, phosphorylation, and Ca2+ binding) will be studied by fluorescence experiments, reaction with site-specific antibodies, and differential scanning calorimetry. The results obtained will be combined in an attempt to produce a functionally relevant structural model for Ca2+-
ATPase.

Funding Scheme

CSC - Cost-sharing contracts

Coordinator

Århus Universitet
Address
185,Ole Worms Allé
8000 Århus C
Denmark

Participants (4)

CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE
France
Address
Cea-saclay
91191 Gif-sur-yvette
CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE
France
Address
Avenue De La Terrasse, Bftiment 26
91198 Gif Sur Yvette
UNIVERSITY OF SOUTHAMPTON
United Kingdom
Address
Bassett Crescent East
SO16 7PX Southampton
Universidad de Extremadura
Spain
Address
Avenida De Elvas
06071 Badajoz