2 Hâ?? 13 C correlation solid-state NMR for investigating dynamics and water accessibilities of proteins and carbohydrates

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2 Hâ?? 13 C correlation solid-state NMR for investigating dynamics and water accessibilities of proteins and carbohydrates

Abstract

Site-specific determination of molecular motion and water accessibility by indirect detection of 2H NMR spectra has advantages over dipolar-coupling based techniques due to the large quadrupolar couplings and the ensuing high angular resolution. Recently, a Rotor Echo Short Pulse IRrAdiaTION mediated cross polarization (RESPIRATIONCP) technique was developed, which allowed efficient transfer of 2H magnetization to 13C at moderate 2H radiofrequency field strengths available on most commercial MAS probes. In this work, we investigate the 2Hâ??13C magnetization transfer characteristics of one-bond perdeuterated CD n spin systems and two-bond H/D exchanged Câ??(O)â??D and Câ??(N)â??D spin systems in carbohydrates and proteins. Our results show that multi-bond, broadband 2Hâ??13C polarization transfer can be achieved using 2H radiofrequency fields of ~50Â*kHz, relatively short contact times of 1.3â??1.7Â*ms, and with sufficiently high sensitivity to enable 2D 2Hâ??13C correlation experiments with undistorted 2H spectra in the indirect dimension. To demonstrate the utility of this 2Hâ??13C technique for studying molecular motion, we show 2Hâ??13C correlation spectra of perdeuterated bacterial cellulose, whose surface glucan chains exhibit a motionally averaged C6 2H quadrupolar coupling that indicates fast trans-gauche isomerization about the C5â??C6 bond. In comparison, the interior chains in the microfibril core are fully immobilized. Application of the 2Hâ??13C correlation experiment to H/D exchanged Arabidopsis primary cell walls show that the Oâ??D quadrupolar spectra of the highest polysaccharide peaks can be fit to a two-component model, in which 74% of the spectral intensity, assigned to cellulose, has a near-rigid-limit coupling, while 26% of the intensity, assigned to matrix polysaccharides, has a weakened coupling of 50Â*kHz. The latter Oâ??D quadrupolar order parameter of 0.22 is significantly smaller than previously reported Câ??D dipolar order parameters of 0.46â??0.55 for pectins, suggesting that additional motions exist at the Câ??O bonds in the wall polysaccharides. 2Hâ??13C polarization transfer profiles are also compared between statistically deuterated and H/D exchanged GB1.



Source: Journal of Biomolecular NMR