Probing and Manipulating Protein Conformation Changes by Time-Resolved Single-Molecule Spectroscopy and Site-Specific Ultramicroscopy
Abstract
This project was to develop and demonstrate a novel single-molecule spectroscopy and imaging technique to support the DARPA program on Control of Protein Conformations. The goal of our project is to advance, integrate, and apply cutting-edge atomic force microscopy (AFM) and single-molecule imaging techniques to control and analyze the protein conformational changes and according activity changes through AFM force manipulation and fluorescence resonant energy transfer (FRET) measurements as a function of local site geometry and molecular structure. In this project, we have focused our demonstration and study on controlling protein conformations to manipulate the enzyme reactivity, affinity, and selectivity. Our approach has been to use AFM tip to control protein conformational states and to apply single-molecule FRET imaging to measure the protein enzyme activity in real time. Our progress towards the demonstration of the new technology has provided an unprecedented advancement on a high spatially (nanometers) and temporally (micro's to seconds) resolved single-molecule spectroscopy of optically and mechanically controlling single-molecule protein conformational changes and activities.
Document Details
- Document Type
- Technical Report
- Publication Date
- Apr 01, 2008
- Accession Number
- ADA500765
Entities
People
- H. P. Lu
Organizations
- Bowling Green State University