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QBE Seminar: ‘Unfolding and Refolding of Individual Bacteriorhodopsin Molecules Probed with 1-µs Resolution’
February 28 @ 1:00 pm - 2:00 pmFree
ABSTRACT: High-precision single-molecule force spectroscopy studies can yield kinetic rate constants, energetics, intermediate states, unfolding pathways, and even a projection of the underlying free-energy landscape. We developed a set of technical advances to atomic force microscopy (AFM) that enabled us to reexamine the unfolding of individual bacteriorhodopsin (bR) molecules embedded in their native lipid bilayer with a 100-fold improvement in time resolution and a 10-fold improvement in force precision. We next integrated these technical advances with site-specific covalent coupling of bR to an AFM tip to quantify the initial unfolding of bR. Hence, we have established a platform for precisely quantifying membrane-protein energetics under native-like conditions. I will conclude by discussing extensions of this capability to other diverse biomolecules, including nucleic-acid structures and globular proteins.
BIO: Thomas Perkins is a Fellow of JILA, a joint institute between NIST and the University of Colorado. He is also acting chief of NIST’s Quantum Physics Division, and a Professor (adjoint) in the Molecular, Cellular and Developmental Biology Department at the University of Colorado. He graduated from Harvard University, did his graduate work at Stanford under Steve Chu, and post-doctoral studies at Princeton and Stanford with Steve Block. Tom is a Fellow of the American Association for the Advancement of Science and the American Physical Society