Science 16 May 2014:
Vol. 344 no. 6185 pp. 742-746
DOI: 10.1126/science.1248167
REPORT
NMR Spectroscopy of Native and in Vitro Tissues Implicates PolyADP Ribose in Biomineralization
W. Ying Chow1, Rakesh Rajan2, Karin H. Muller3, David G. Reid1, Jeremy N. Skepper3, Wai Ching Wong1, Roger A. Brooks2, Maggie Green4, Dominique Bihan5, Richard W. Farndale5, David A. Slatter5, Catherine M. Shanahan6, Melinda J. Duer1,*
+ Author Affiliations
1Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK.
2Orthopaedic Research Unit, University of Cambridge, Addenbrooke’s Hospital, Cambridge CB2 0QQ, UK.
3Department of Physiology, Development, and Neuroscience, University of Cambridge, Downing Site, Cambridge CB2 3DY, UK.
4Central Biomedical Resources, University of Cambridge, School of Clinical Medicine, West Forvie Building, Forvie Site, Robinson Way, Cambridge CB2 0SZ, UK.
5Department of Biochemistry, University of Cambridge, Downing Site, Cambridge CB2 1QW, UK.
6British Heart Foundation Centre of Research Excellence, Cardiovascular Division, James Black Centre, King’s College London, 125 Coldharbour Lane, London SE5 9NU, UK
?*Corresponding author. E-mail: mjd13@cam.ac.uk
ABSTRACTEDITOR'S SUMMARY
Nuclear magnetic resonance (NMR) spectroscopy is useful to determine molecular structure in tissues grown in vitro only if their fidelity, relative to native tissue, can be established. Here, we use multidimensional NMR spectra of animal and in vitro model tissues as fingerprints of their respective molecular structures, allowing us to compare the intact tissues at atomic length scales. To obtain spectra from animal tissues, we developed a heavy mouse enriched by about 20% in the NMR-active isotopes carbon-13 and nitrogen-15. The resulting spectra allowed us to refine an in vitro model of developing bone and to probe its detailed structure. The identification of an unexpected molecule, poly(adenosine diphosphate ribose), that may be implicated in calcification of the bone matrix, illustrates the analytical power of this approach.