Full-length Vpu and human CD4(372-433) in phospholipid bilayers as seen by magic angle spinning NMR.
Related Articles Full-length Vpu and human CD4(372-433) in phospholipid bilayers as seen by magic angle spinning NMR.
Biol Chem. 2013 Jul 17;
Authors: Do HQ, Wittlich M, Glück JM, Möckel L, Willbold D, Koenig BW, Heise H
Abstract
Abstract HIV-1 Vpu and CD4(372-433), a peptide comprising the transmembrane and cytoplasmic domain of human CD4, were recombinantly expressed in Escherichia coli, uniformly labeled with 13C und 15N isotopes, and separately reconstituted into phospholipid bilayers. Highly resolved, dipolar cross polarization-based solid-state NMR spectra of the two transmembrane proteins were recorded under magic angle sample spinning (MAS). Partial assignment of 13C resonances was achieved. Site-specific assignments were obtained for 13 amino acid residues of CD4(372-433) and two VpU residues. Additional amino acid type-specific assignments were achieved for 10 amino acid spin systems for both CD4(372-433) and Vpu. Further, structural flexibility was probed with different dipolar recoupling techniques, and the correct insertion of the TM domains into the lipid bilayers was confirmed by proton spin diffusion experiments.
PMID: 23863698 [PubMed - as supplied by publisher]
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