Plant cell-wall cross-links by REDOR NMR spectroscopy |
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Authors: | Cegelski Lynette O'Connor Robert D Stueber Dirk Singh Manmilan Poliks Barbara Schaefer Jacob |
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Institution: | Department of Chemistry, Washington University, St. Louis, Missouri 63130, United States. cegelski@stanford.edu |
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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. |
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