Dipolar Assisted Assignment Protocol (DAAP) for MAS solid-state NMR of rotationally aligned membrane proteins in phospholipid bilayers.
A method for making resonance assignments in magic angle spinning solid-state NMR spectra of membrane proteins that utilizes the range of heteronuclear dipolar coupling frequencies in combination with conventional chemical shift based assignment methods is demonstrated. The Dipolar Assisted Assignment Protocol (DAAP) takes advantage of the rotational alignment of the membrane proteins in liquid crystalline phospholipid bilayers. Improved resolution is obtained by combining the magnetically inequivalent heteronuclear dipolar frequencies with isotropic chemical shift frequencies. Spectra with both dipolar and chemical shift frequency axes assist with resonance assignments. DAAP can be readily extended to three- and four-dimensional experiments and to include both backbone and side chain sites in proteins.
Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA 92093-0307, United States.
Bibhuti B
BB
Das
Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA 92093-0307, United States.
Hua
H
Zhang
Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA 92093-0307, United States. Electronic address: sopella@ucsd.edu.
Stanley J
SJ
Opella
D000465:Algorithms; D008051:Lipid Bilayers / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry*; D010743:Phospholipids / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D008051:Lipid Bilayers; D008565:Membrane Proteins; D010743:Phospholipids
MAS; Membrane protein; Solid state NMR; Vpu