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Biophysical Journal 64: 614-621 (1993)
© 1993 the Biophysical Society

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Measurement of rotational dynamics by the simultaneous nonlinear analysis of optical and EPR data.

E J Hustedt, C E Cobb, A H Beth and J M Beechem

Vanderbilt University, Nashville, Tennessee 37232.

ABSTRACT

In the preceding companion article in this issue, an optical dye and a nitroxide radical were combined in a new dual function probe, 5-SLE. In this report, it is demonstrated that time-resolved optical anisotropy and electron paramagnetic resonance (EPR) data can be combined in a single analysis to measure rotational dynamics. Rigid-limit and rotational diffusion models for simulating nitroxide EPR data have been incorporated into a general non-linear least-squares procedure based on the Marquardt-Levenberg algorithm. Simultaneous fits to simulated time-resolved fluorescence anisotropy and linear EPR data, together with simultaneous fits to experimental time-resolved phosphorescence anisotropy decays and saturation transfer EPR (ST-EPR) spectra of 5-SLE noncovalently bound to bovine serum albumin (BSA) have been performed. These results demonstrate that data from optical and EPR experiments can be combined and globally fit to a single dynamic model.




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Copyright © 1993 by the Biophysical Society.