Festkörper‐NMR‐Spektroskopie an komplexen Biomolekülen

Abstract

Biomolekulare Anwendungen der NMR‐Spektroskopie werden oft mit löslichen Molekülen oder Magnetresonanztomographie in Verbindung gebracht. Seit Ende der 70er Jahre liefert darüber hinaus die Festkörper‐NMR‐Spektroskopie (FK‐NMR) auf atomarer Ebene Einblicke in komplexe biomolekulare Systeme von Lipid‐Doppelschichten bis hin zu Biomaterialien. Im letzten Jahrzehnt haben Fortschritte im Bereich der NMR‐Spektroskopie, der Biophysik und der Molekularbiologie das Repertoire der FK‐NMR‐Spektroskopie für biomolekulare Studien erheblich erweitert. Dieser Aufsatz behandelt neueste Ansätze und deren methodische Herausforderungen und diskutiert Fortschritte bei der Anwendung der FK‐NMR‐Spektroskopie im Grenzgebiet von Struktur‐ und Zellbiologie.

Dual Acquisition Magic‐Angle Spinning Solid‐State NMR‐Spectroscopy: Simultaneous Acquisition of Multidimensional Spectra of Biomacromolecules

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Magic-angle spinning solid-state NMR (MAS ssNMR) spectroscopy is a powerful method for structure determination of biomacromolecules that are recalcitrant to crystallization (membrane proteins and fibrils). [1,[2][3][4] Developments in pulse sequence design, [5][6][7][8][9][10] probes, [11] isotopic labeling schemes, [12] sensitivity enhancement using paramagnetic effects, [13] dynamic nuclear polarization (DNP), [14][15][16] and sample preparations have made it possible to advance resonance assignments and structure determination by ssNMR spectroscopy. However, as the biological systems under investigation increase in complexity, new methods are needed to improve spectral resolution and sensitivity as well as to speed up NMR data acquisition.Here we present a novel approach (which we refer to it as "dual" acquisition MAS ssNMR or DUMAS ssNMR spectroscopy) for parallel acquisition of multidimensional ssNMR experiments without the need of additional hardware.

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Combination of DQ and ZQ Coherences for Sensitive Through‐Bond NMR Correlation Experiments in Biosolids under Ultra‐Fast MAS

“…The transverse magnetization is present on spin S during the second refocusing echo of the sequences, and therefore the signal detected decays during the second echo delay D 2 according to the transverse dephasing times T 2 ' S of spin S, following summing the result. (2)]. Traces parallel to w 2 taken from the DQ-SQ and ZQ-SQ spectra are shown as insets in (a), while traces through the two sheared spectra and resulting summed spectrum can be seen as insets in (b).…”

“…[1,2] The recent introduction of small diameter rotors (1.3 mm), capable of spinning the sample at frequencies up to 60 kHz [ultra-fast magic angle spinning (MAS)], has opened new avenues in this area, such as, to name but a few, accelerated acquisition, [3][4][5][6] the feasibility of exploiting proton-proton distance restraints in three-dimensional structure calculations, [7] or the possibility of probing site specifically sub-nanosecond molecular motions using novel dynamics probes. Steadily ongoing methodological developments combined with tremendous engineering advances in probe and spectrometer hardware, and notably increased magnetic field strengths have paved the way for studying structure and dynamics of solid chemical and biological samples at atomic resolution, spanning a broad atlas of structures ranging from materials to protein aggregates or membrane proteins.…”

“…Steadily ongoing methodological developments combined with tremendous engineering advances in probe and spectrometer hardware, and notably increased magnetic field strengths have paved the way for studying structure and dynamics of solid chemical and biological samples at atomic resolution, spanning a broad atlas of structures ranging from materials to protein aggregates or membrane proteins. [1,2] The recent introduction of small diameter rotors (1.3 mm), capable of spinning the sample at frequencies up to 60 kHz [ultra-fast magic angle spinning (MAS)], has opened new avenues in this area, such as, to name but a few, accelerated acquisition, [3][4][5][6] the feasibility of exploiting proton-proton distance restraints in three-dimensional structure calculations, [7] or the possibility of probing site specifically sub-nanosecond molecular motions using novel dynamics probes. [8] Like in solution, the assignment of resonances is a prerequisite for any structural or dynamics investigation carried out by solid-state NMR spectroscopy.…”

Conformation and Topology of Diacylglycerol Kinase in E.coli Membranes Revealed by Solid‐state NMR Spectroscopy

“…Magic angle spinning (MAS) solid-state NMR (ssNMR) is a powerful tool for studying the structures of membrane Figure S4). proteins [9][10][11] directly within lipid bilayers [12][13][14][15][16][17][18] and even within whole cells. [19,20] In this study, 3D MAS NMR was employed to characterize the structure of DAGK reconstituted into its native E.coli lipid membranes.…”