Simultaneous acquisition of 2D and 3D solid-state NMR experiments for sequential assignment of oriented membrane protein samples

Gopinath Tata, Kaustubh R. Mote, Gianluigi Veglia

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

Abstract

Abstract We present a new method called DAISY (Dual Acquisition orIented ssNMR spectroScopY) for the simultaneous acquisition of 2D and 3D oriented solid-state NMR experiments for membrane proteins reconstituted in mechanically or magnetically aligned lipid bilayers. DAISY utilizes dual acquisition of sine and cosine dipolar or chemical shift coherences and long living 15N longitudinal polarization to obtain two multi-dimensional spectra, simultaneously. In these new experiments, the first acquisition gives the polarization inversion spin exchange at the magic angle (PISEMA) or heteronuclear correlation (HETCOR) spectra, the second acquisition gives PISEMA-mixing or HETCOR-mixing spectra, where the mixing element enables inter-residue correlations through 15N-15N homonuclear polarization transfer. The analysis of the two 2D spectra (first and second acquisitions) enables one to distinguish 15N-15N inter-residue correlations for sequential assignment of membrane proteins. DAISY can be implemented in 3D experiments that include the polarization inversion spin exchange at magic angle via I spin coherence (PISEMAI) sequence, as we show for the simultaneous acquisition of 3D PISEMAI-HETCOR and 3D PISEMAI-HETCOR-mixing experiments. 2015

Original languageEnglish (US)
Article number9916
Pages (from-to)53-61
Number of pages9
JournalJournal of biomolecular NMR
Volume62
Issue number1
DOIs
StatePublished - May 1 2015

Keywords

  • Aligned lipid bilayers
  • Bicelles
  • Membrane proteins
  • Oriented solid-state NMR
  • Simultaneous acquisition

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