Mary Barkley

Professor

Ph.D. University of California,1969
View Curriculum Vitae

Structure and Dynamics of Biological Macromolecule

RESEARCH DESCRIPTION

Our research group investigates the structure and dynamics of biological macromolecules. With the completion of the human genome project, attention is turning to the proteins that are encoded by the genome. Better understanding of how protein structure dictates protein function makes it possible to design better drugs to combat disease. Viral replication is a key target for drug therapy. We are currently studying enzymes from two viruses: the AIDS virus HIV-1 and hepatitis C virus HCV.

Our laboratory is using fluorescence spectroscopy and other biophysical techniques to understand how protein structure affects protein function. After a molecule absorbs a photon of light, a few billionths of a second later it emits a photon of visible light called fluorescence. The fluorescent light becomes a probe to follow changes in protein structure that are important for protein function. We attach fluorescent probes to engineered proteins and oligonucleotides. We also study intrinsic tryptophan fluorescence as a probe of protein structure. We use pre-steady-state kinetics to study enzyme reaction mechanisms and analytical ultracentrifugation to study protein-protein and protein-nucleic acid interactions.

Reverse transcriptase catalyzes the first step in HIV replication by converting single-stranded viral RNA into double-stranded proviral DNA. We are studying the conformational changes in reverse transcriptase upon binding of RNA and DNA substrates and inhibitors using pre-steady state kinetics and fluorescence techniques. Heterodimer formation is examined by analytical ultracentrifugation. These experiments involve site-specific fluorescent labeling of reverse transcriptase as well as RNA and DNA substrates containing fluorescent base analogs. NS5B RNA polymerase synthesizes HCV viral RNA, but may require other viral proteins and also host cell proteins. We are studying the interactions of NS5B polymerase with RNA and other viral enzymes by fluorescence spectroscopy and analytical ultracentrifugation.

RELATED RESEARCH AREAS

View Mary Barkley's Publications on PubMed