Automated Assignment in Selectively Methyl-Labeled Proteins

Yingqi Xu, Minhao Liu, Peter J. Simpson, Rivka Isaacson, Ernesto Cota, Jan Marchant, Daiwen Yang, Xiaodong Zhang, Paul Freemont, Stephen Matthews*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

29 Citations (Scopus)

Abstract

Specific methyl labeling schemes and transverse relaxation optimized spectroscopy (TROSY) has extended the molecular size range for the application of NMR spectroscopy to very large proteins (up to similar to 1 MDa). Existing strategies for resonance assignment of methyl groups in large systems are based on NMR spectra recorded on smaller fragments and mutants. This is very time-consuming, and chemical shift changes due to mutation or truncation can often complicate interpretation. We have developed a new automated procedure able to rapidly assign the majority of methyl. groups in very large proteins, without recourse to mutagenesis or truncated fragments (http://nmr.bc.ic.ac.uk/map-xs/). We demonstrate the effectiveness of this approach on the 300 kDa, ILV-labeled proteasome (alpha(7)alpha(7)) for which excellent spectra have been previously recorded. Of the observed methyl groups, 99% can be correctly assigned in a matter of minutes without manual intervention.

Original languageEnglish
Pages (from-to)9480-9481
Number of pages1
JournalJournal of the American Chemical Society
Volume131
Issue number27
DOIs
Publication statusPublished - 15 Jul 2009

Keywords

  • RELAXATION
  • NMR-SPECTROSCOPY
  • ALANINE
  • CHEMICAL-SHIFTS
  • MOLECULAR-WEIGHT PROTEINS
  • COMPLEXES

Fingerprint

Dive into the research topics of 'Automated Assignment in Selectively Methyl-Labeled Proteins'. Together they form a unique fingerprint.

Cite this