NMR spectroscopy of native and in vitro tissues implicates polyADP ribose in biomineralization.
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.
Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK.
W Ying
WY
Chow
Orthopaedic Research Unit, University of Cambridge, Addenbrooke's Hospital, Cambridge CB2 0QQ, UK.
Rakesh
R
Rajan
Department of Physiology, Development, and Neuroscience, University of Cambridge, Downing Site, Cambridge CB2 3DY, UK.
Karin H
KH
Muller
Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK.
David G
DG
Reid
Department of Physiology, Development, and Neuroscience, University of Cambridge, Downing Site, Cambridge CB2 3DY, UK.
Jeremy N
JN
Skepper
Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK.
Wai Ching
WC
Wong
Orthopaedic Research Unit, University of Cambridge, Addenbrooke's Hospital, Cambridge CB2 0QQ, UK.
Roger A
RA
Brooks
Central Biomedical Resources, University of Cambridge, School of Clinical Medicine, West Forvie Building, Forvie Site, Robinson Way, Cambridge CB2 0SZ, UK.
Maggie
M
Green
Department of Biochemistry, University of Cambridge, Downing Site, Cambridge CB2 1QW, UK.
Dominique
D
Bihan
Department of Biochemistry, University of Cambridge, Downing Site, Cambridge CB2 1QW, UK.
Richard W
RW
Farndale
Department of Biochemistry, University of Cambridge, Downing Site, Cambridge CB2 1QW, UK.
David A
DA
Slatter
British Heart Foundation Centre of Research Excellence, Cardiovascular Division, James Black Centre, King's College London, 125 Coldharbour Lane, London SE5 9NU, UK.
Catherine M
CM
Shanahan
Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK. mjd13@cam.ac.uk.
Melinda J
MJ
Duer
D000818:Animals; D001846:Bone Development; D002113:Calcification, Physiologic; D002247:Carbon Isotopes; D005109:Extracellular Matrix / Q000737:chemistry; D006132:Growth Plate / Q000254:growth & development; D051379:Mice; D008954:Models, Biological; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011064:Poly Adenosine Diphosphate Ribose / Q000032:analysis*; D012756:Sheep
D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D011064:Poly Adenosine Diphosphate Ribose