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The affinity and structure of biomolecules immobilized on a 3-D substrate: Time-resolved EPR detection during mobile phase gradients


EMSL Project ID
39731

Abstract

We propose the development of a new magnetic resonance technique to monitor the interactions of biomolecules with binding partners and their environment. Physically, this will be implemented by designing a fritted column to replace the sample holder in the EPR spectrometer. This column will be loaded with biomolecules tethered to a resin substrate. A mobile phase will be flowed through the column and changes in the EPR signal monitored. These changes can reflect a number of different processes, depending on experiment design. For instance, the affinity of the tethered molecule for a paramagnetic species in the mobile phase will be reflected in the increasing EPR signal with time. Similarly, if the tethered molecule has an EPR spectrum that is sensitive to structural changes, the stability of the tethered molecule can be determined by a mobile phase gradient of increasing chaotrope concentration.

Project Details

Start Date
2010-04-01
End Date
2012-11-07
Status
Closed

Team

Principal Investigator

Eric Walter
Institution
Environmental Molecular Sciences Laboratory

Team Members

Eric Evans
Institution
University of California, Santa Cruz

Mary Cloninger
Institution
Montana State University

Romulus Turcu
Institution
Babes-Bolyai University

Glenn Millhauser
Institution
University of California, Santa Cruz

Uma Aryal
Institution
Pacific Northwest National Laboratory

Andrew Lipton
Institution
Environmental Molecular Sciences Laboratory

Related Publications

Dudzik CG, ED Walter, BS Abrams, MS Jurica, and GL Millhauser. 2013. "Coordination of Copper to the Membrane-Bound Form of ?-Synuclein." Biochemistry 52(1):53-60. doi:10.1021/bi301475q