[2,3-C-13]-labeling of aromatic residues-getting a head start in the magic-angle-spinning NMR assignment of membrane proteins

Matthias Hiller, Victoria A. Higman, Stefan Jehle, Barth-Jan van Rossum, Werner Kuehlbrandt, Hartmut Oschkinat*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

40 Citations (Scopus)

Abstract

The assignment of magic-angle spining NMR speara of large membrane proteins such as the 281-residue ouler membrane protein G (OmpG) is difficult because of substantial signal overlap and line-broadening effects. To obtain sequence specific assignments a new labeling sitalegy was applied to OmpG in which alanine and glycine were uniformly (N-15, C-13] labeled and phenylalanine and tyrosine were [N-15, C alpha,C beta-C-13]-labeled. This labeling pattern resulted in many long-range inter-residue cross- peaks being observed in spectra recorded with long mixing times. In addition, a reduction in the number of C' labels resulted in NCO-type spectra with little overlap and provided the basis for unambiguous sequential assignments. In conjuntion with spectra from [1,3-C-13]. and [2-C-13]-glycerol samples, a total of 45 reliable sequence specific assignments could be made for OmpG on the basis of this sample.

Original languageEnglish
Pages (from-to)408-+
Number of pages3
JournalJournal of the American Chemical Society
Volume130
Issue number2
DOIs
Publication statusPublished - 16 Jan 2008

Keywords

  • UBIQUITIN
  • SPECTROSCOPY
  • RECEPTOR
  • PORIN PROTEIN
  • C-13
  • OMPG
  • ESCHERICHIA-COLI K-12
  • MAS
  • COIL
  • SOLID-STATE NMR

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