Determination of Membrane Protein Structure and Dynamics by Magic-Angle-Spinning Solid-State NMR Spectroscopy

• Published in: Journal of the American Chemical Society Vol. 127; no. 37; pp. 12965 - 12974
• Main Authors: Andronesi, Ovidiu C, Becker, Stefan, Seidel, Karsten, Heise, Henrike, Young, Howard S, Baldus, Marc
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 21-09-2005
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Calcium-Binding Proteins - chemistry; Carbon Isotopes; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Molecular biophysics; Nitrogen Isotopes; Nuclear Magnetic Resonance, Biomolecular - methods; Protein Structure, Secondary; Proteins; Sensitivity and Specificity; Spectroscopy : techniques and spectras; Spin Labels; Thermodynamics; Solid state; Membrane protein; Biochemical analysis; Nitrogen 15; NMR spectrometry; Carbon 13; Labelled compound; Heteronuclear correlation; Structural analysis; Two dimension spectrometry; Pulse sequence
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/ja0530164

It is shown that molecular structure and dynamics of a uniformly labeled membrane protein can be studied under magic-angle-spinning conditions. For this purpose, dipolar recoupling experiments are combined with novel through-bond correlation schemes that probe mobile protein segments. These NMR schemes are demonstrated on a uniformly [13C,15N] variant of the 52-residue polypeptide phospholamban. When reconstituted in lipid bilayers, the NMR data are consistent with an α-helical trans-membrane segment and a cytoplasmic domain that exhibits a high degree of structural disorder.