Plant cell-wall cross-links by REDOR NMR spectroscopy

J Am Chem Soc. 2010 Nov 17;132(45):16052-7. doi: 10.1021/ja104827k. Epub 2010 Oct 21.

Abstract

We present a new method that integrates selective biosynthetic labeling and solid-state NMR detection to identify in situ important protein cross-links in plant cell walls. We have labeled soybean cells by growth in media containing l-[ring-d(4)]tyrosine and l-[ring-4-(13)C]tyrosine, compared whole-cell and cell-wall (13)C CPMAS spectra, and examined intact cell walls using (13)C{(2)H} rotational echo double-resonance (REDOR) solid-state NMR. The proximity of (13)C and (2)H labels shows that 25% of the tyrosines in soybean cell walls are part of isodityrosine cross-links between protein chains. We also used (15)N{(13)C} REDOR of intact cell walls labeled by l-[ε-(15)N,6-(13)C]lysine and depleted in natural-abundance (15)N to establish that the side chains of lysine are not significantly involved in covalent cross-links to proteins or sugars.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Cell Wall / chemistry*
  • Cells, Cultured
  • Glycine max / chemistry
  • Glycine max / cytology
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Plant Proteins / analysis*
  • Tyrosine / analogs & derivatives*
  • Tyrosine / analysis

Substances

  • Plant Proteins
  • Tyrosine
  • isodityrosine