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4D APSY-HBCB(CG)CDHD experiment for automated assignment of aromatic amino acid side chains in proteins


Abstract A four-dimensional (4D) APSY (automated projection spectroscopy)-HBCB(CG)CDHD experiment is presented. This 4D experiment correlates aromatic with aliphatic carbon and proton resonances from the same amino acid side chain of proteins in aqueous solution. It thus allows unambiguous sequence-specific assignment of aromatic amino acid ring signals based on backbone assignments. Compared to conventional 2D approaches, the inclusion of evolution periods on 1Hβ and 13Cδ efficiently removes overlaps, and provides two additional frequencies for consequent automated or manual matching. The experiment was successfully applied to three proteins with molecular weights from 6 to 13 kDa. For the complementation of the assignment of the aromatic resonances, TOCSY- or COSY-based versions of a 4D APSY-HCCHaro sequence are proposed.

• Content Type Journal Article
• Category Article
• Pages 1-6
• DOI 10.1007/s10858-011-9572-7
• Authors
  • Barbara Krähenbühl, Institute of Molecular Biology and Biophysics, ETH Zurich, 8093 Zurich, Switzerland
  • Sebastian Hiller, Biozentrum, University of Basel, 4056 Basel, Switzerland
  • Gerhard Wider, Institute of Molecular Biology and Biophysics, ETH Zurich, 8093 Zurich, Switzerland

• • Journal Journal of Biomolecular NMR
  • Online ISSN 1573-5001
  • Print ISSN 0925-2738

Source: Journal of Biomolecular NMR