Objective
This joint project will explore the molecular mechanism of one of the fundamental problems in biological science: the transduction of chemical energy into mechanical work performed by motor proteins. The main goal of the proposed research is to understand how the conformational changes taking place in the myosin head during ATP binding and hydrolysis are connected with transition of the head between different acting-binding states ("weak" and "strong" binding states). For this purpose stable analogues of myosin ATPase intermediates (the ternary complexes of the myosin head with ADP and different phosphate analogues such as orthovanadate, beryllium fluoride, or aluminium fluoride) will be used. Information on structural changes in the myosin head induced by the formation of these complexes will be obtained by studying the thermal denaturation of the myosin head by using of the method of differential scanning calorimetry.
The interaction of these complexes with acting in the absence and in the presence of regulatory proteins (tropomyosin & troponin) will be studied by stopped-flow kinetic measurements as well as by isothermal titration microcalorimetry. Then, site-directed mutagenesis of the myosin head (using Dictyostelium discoideum myosin) will be used to reveal the sites in the head important for the information transfer through the head structure, from the ATPase site to actin-binding sites. The results of these experiments will provide new insights into molecular mechanism of energy transduction in the force generation process.
Topic(s)
Call for proposal
Data not availableFunding Scheme
Data not availableCoordinator
CANTERBURY
United Kingdom