A pressure-jump EPR system to monitor millisecond conformational exchange rates of spin-labeled proteins.

ABSTRACT: Site-directed spin labeling electron paramagnetic resonance (SDSL-EPR) using nitroxide spin labels is a well-established technology for mapping site-specific secondary and tertiary structure and for monitoring conformational changes in proteins of any degree of complexity, including membrane proteins, with high sensitivity. SDSL-EPR also provides information on protein dynamics in the time scale of ps-µs using continuous wave lineshape analysis and spin lattice relaxation time methods. However, the functionally important time domain of µs-ms, corresponding to large-scale protein motions, is inaccessible to those methods. To extend SDSL-EPR to the longer time domain, the perturbation method of pressure-jump relaxation is implemented. Here, we describe a complete high-pressure EPR system at

SUBMITTER: Grosskopf JD 

PROVIDER: S-EPMC11100676 | biostudies-literature | 2024 May

REPOSITORIES: biostudies-literature