Award details

Single molecule investigations of the mechanical chemical and structural properties of biomolecules

Reference	BB/D525013/1
Principal Investigator / Supervisor	Professor David Brockwell
Co-Investigators / Co-Supervisors	Professor Stephen Evans, Professor Sheena Radford, Professor David Smith, Dr Neil Thomson
Institution	University of Leeds
Department	Inst of Molecular & Cellular Biology
Funding type	Research
Value (£)	94,400
Status	Completed
Type	Research Grant
Start date	01/02/2006
End date	31/10/2006
Duration	9 months

Abstract

This proposal seeks funds to purchase an Asylum MFP-3D, a bioheater and active vibration damping platform. Non-covalent biomolecular interactions play an important role in many phenomena in biology such as protein folding, bilayer formation (phase separation), protein-ligand interactions and protein-protein interactions in biomolecular assembly. All of these processes occur at a length scale on which the imaging capability of the Atomic Force Microscope (AFM) can yield structural information. The development of force spectroscopy, the ability to calculate the strength of interaction by measuring the force at which a `bond¿ ruptures, has transformed our understanding of these processes on the single molecule level and allows detailed investigation of the factors controlling these complex phenomena. Previously, in order to perform single molecule force spectroscopy with adequate signal to noise and stability, it has been necessary to use specially designed AFMs that have no imaging facility. However, the Asylum MFP-3D is both a state of the art force spectroscope and a state of the art imaging AFM that can image over a very large area. This instrument, therefore, not only allows previously impossible experiments to be performed by will also provide an additional force probe ¿ access to our current instrument is now causing a severe bottleneck in several areas of research. Additionally, purchase of a bioheater, which is packaged with a closed fluid cell, allows temperature control and solvent exchange and also reduces solute evaporation ¿ a major drawback of the MFP-1D. The provision of an Asylum MFP-3D will allow: 1. protein mechanical unfolding data to be more rapidly and efficiently accumulated and will also allow the effects of temperature to be assessed on the mechanical properties of proteins. 2. the development of novel tethering strategies for in vitro lipid bilayers to facilitate the study of membrane proteins. 3. the investigation of cytoskeleton assembly forapplication in engineered biocompatible surfaces. 4. the characterisation of the spatial and functional relationships between DNA and processive enzymes. 5. the determination of the mechanical response of amyloid fibrils using a range of force measurements and the exploration of the sequence dependence of the mechanical response. 6. the characterisation of size, microstructure and dynamics of lipid rafts and their role in processes such as signal transduction and membrane trafficking.

Summary

unavailable

Committee	Closed Committee - Biomolecular Sciences (BMS)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	Research Equipment Initiative 2005 (RE5) [2005]
Funding Scheme	X – not Funded via a specific Funding Scheme

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