Stationary-phase EPR for exploring protein structure, conformation, and dynamics in spin-labeled proteins.

ABSTRACT: Proteins tethered to solid supports are of increasing interest in bioanalytical chemistry and protein science in general. However, the extent to which tethering modifies the energy landscape and dynamics of the protein is most often unknown because there are few biophysical methods that can determine secondary and tertiary structures and explore conformational equilibria and dynamics of a tethered protein with site-specific resolution. Site-directed spin labeling (SDSL) combined with electron paramagnetic resonance (EPR) offers a unique opportunity for this purpose. Here, we employ a general strategy using unnatural amino acids that enables efficient and site-specific tethering of a spin-labeled protein to a Sepharose solid support. Remarkably, EPR spectra of spin-labeled T4 lysozyme (T4L)

SUBMITTER: Lopez CJ 

PROVIDER: S-EPMC4238802 | biostudies-literature | 2014 Nov

REPOSITORIES: biostudies-literature