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Optimal isotope labelling for NMR protein structure determinations

Author

Listed:
  • Masatsune Kainosho

    (Tokyo Metropolitan University)

  • Takuya Torizawa

    (Tokyo Metropolitan University)

  • Yuki Iwashita

    (Tokyo Metropolitan University)

  • Tsutomu Terauchi

    (Tokyo Metropolitan University)

  • Akira Mei Ono

    (Tokyo Metropolitan University)

  • Peter Güntert

    (RIKEN Genomic Sciences Center)

Abstract

Nuclear-magnetic-resonance spectroscopy can determine the three-dimensional structure of proteins in solution. However, its potential has been limited by the difficulty of interpreting NMR spectra in the presence of broadened and overlapping resonance lines and low signal-to-noise ratios. Here we present stereo-array isotope labelling (SAIL), a technique that can overcome many of these problems by applying a complete stereospecific and regiospecific pattern of stable isotopes that is optimal with regard to the quality and information content of the resulting NMR spectra. SAIL uses exclusively chemically and enzymatically synthesized amino acids for cell-free protein expression. We demonstrate for the 17-kDa protein calmodulin and the 41-kDa maltodextrin-binding protein that SAIL offers sharpened lines, spectral simplification without loss of information, and the ability to rapidly collect the structural restraints required to solve a high-quality solution structure for proteins twice as large as commonly solved by NMR. It thus makes a large class of proteins newly accessible to detailed solution structure determination.

Suggested Citation

  • Masatsune Kainosho & Takuya Torizawa & Yuki Iwashita & Tsutomu Terauchi & Akira Mei Ono & Peter Güntert, 2006. "Optimal isotope labelling for NMR protein structure determinations," Nature, Nature, vol. 440(7080), pages 52-57, March.
    • Handle: RePEc:nat:nature:v:440:y:2006:i:7080:d:10.1038_nature04525
    DOI: 10.1038/nature04525
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    Cited by:

    1. Arash Bahrami & Amir H Assadi & John L Markley & Hamid R Eghbalnia, 2009. "Probabilistic Interaction Network of Evidence Algorithm and its Application to Complete Labeling of Peak Lists from Protein NMR Spectroscopy," PLOS Computational Biology, Public Library of Science, vol. 5(3), pages 1-15, March.

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