Objective Single molecule methods have shed light on many important biochemical phenomena: optical tweezers has contributed important mechanical information about molecular motors, while single molecule imaging techniques have been used to explore the diversity of behaviour of biological molecules which is often masked by ensemble measurements.The central enzyme in the DNA replication process is DNA polymerase, a particularly interesting enzyme responsible for duplicating our genetic information, making less than one mistake in a billion bases. One goal of this research proposal is to develop a method of studying the complex kinetics of DNA polymerase at the single molecule level, using fluorescence microscopy and fluorescently labelled nucleotides.DNA polymerase can be considered a molecular motor since it moves along a DNA track while using energy. Laser tweezers can probe biological systems as a function of force. By studying the enzyme reaction as a function of force, information about the energy release can be obtained.We propose experimental configurations whereby DNA polymerase is immobilized on a solid surface, allowing direct measurement of the interaction forces. Also the coupling of single-molecule manipulation with single-molecule fluorescence opens up many possibilities for observing spectroscopic signals in response to mechanically induced changes in biological macromolecules.This is an ambitious project that initially will allow us to correlate conformational changes and displacement with polymerization or digestion of DNA polymerase The Work Programme contains a series of specific objectives, the augmentative goal of which is to direct attention and funding towards the long-term reinforcement of European science and technology.The interdisciplinary nature of the current proposal as well as the novel concepts and approaches that will be implemented, have been conceived to support and contribute to the achievement of the above goals. Fields of science natural sciencesbiological sciencesgeneticsDNAnatural sciencesphysical sciencesopticsspectroscopyemission spectroscopynatural sciencesphysical sciencesopticsmicroscopynatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymesnatural sciencesphysical sciencesopticslaser physics Keywords DNA polymerase enzyme kinetics force spectroscopy mechanochemistry optical tweezers single molecule fluorescence spectroscopy Programme(s) FP6-MOBILITY - Human resources and Mobility in the specific programme for research, technological development and demonstration "Structuring the European Research Area" under the Sixth Framework Programme 2002-2006 Topic(s) MOBILITY-2.2 - Marie Curie Outgoing International Fellowships (OIF) Call for proposal FP6-2005-MOBILITY-6 See other projects for this call Funding Scheme OIF - Marie Curie actions-Outgoing International Fellowships Coordinator UNIVERSITY COURT OF THE UNIVERSITY OF ST ANDREWS EU contribution No data Address College Gate, North Street ST. ANDREWS United Kingdom See on map Links Website Opens in new window Total cost No data Participants (1) Sort alphabetically Sort by EU Contribution Expand all Collapse all BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIVERSITY United States EU contribution No data Address 310 Panama Street STANFORD, CA See on map Links Website Opens in new window Total cost No data
