Comparison of electron paramagnetic resonance methods to determine distances between spin labels on human carbonic anhydrase II |
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| Authors: | Persson M Harbridge J R Hammarström P Mitri R Mårtensson L G Carlsson U Eaton G R Eaton S S |
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| Affiliation: | Department of Chemistry and Biochemistry, University of Denver, Denver, Colorado 80208, USA. |
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| Abstract: | Four doubly spin-labeled variants of human carbonic anhydrase II and corresponding singly labeled variants were prepared by site-directed spin labeling. The distances between the spin labels were obtained from continuous-wave electron paramagnetic resonance spectra by analysis of the relative intensity of the half-field transition, Fourier deconvolution of line-shape broadening, and computer simulation of line-shape changes. Distances also were determined by four-pulse double electron-electron resonance. For each variant, at least two methods were applicable and reasonable agreement between methods was obtained. Distances ranged from 7 to 24 A. The doubly spin-labeled samples contained some singly labeled protein due to incomplete labeling. The sensitivity of each of the distance determination methods to the non-interacting component was compared. |
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