Polychromatic selective population inversion for TROSY experiments with large proteins |
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Authors: | Bromek Krystyna Lee Donghan Hauhart Richard Krych-Goldberg Malgorzata Atkinson John P Barlow Paul N Pervushin Konstantin |
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Institution: | Laboratory of Physical Chemistry, Swiss Federal Institute of Technology, ETH-H?nggerberg, CH-8093 Zürich, Switzerland. |
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Abstract: | This paper presents polychromatic selective polarization inversion (PC-SPI) as an alternative to the polarization transfer methods recently developed for the application of NMR to large biological molecules. Theoretical and numerical considerations indicate that PC-SPI has the potential for more efficient polarization transfer under conditions of rapid transverse relaxation compared to J coupling- and cross-correlated relaxation-based transfers. The main advantage offered by the method presented here is the maintenance of near-optimal trajectories of inversion of the individual components of the spin magnetization while using broadband optimized pulses. A 2D experiment was implemented combining PC-SPI with TROSY-based chemical shift correlation. The experiment was applied to detect (15)N-(1)H chemical shift correlation spectra of a 200 kDa complex consisting of an 80% (2)H- and uniformly (15)N,(13)C-labeled 22 kDa portion of complement receptor type 1 and unlabeled C3b of complement (180 kDa). |
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