Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK.
Orthopaedic Research Unit, University of Cambridge, Addenbrooke's Hospital, Cambridge CB2 0QQ, UK.
Science. 2014 May 16;344(6185):742-6. doi: 10.1126/science.1248167.
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.
核磁共振(NMR)光谱学在确定体外培养组织的分子结构方面很有用,前提是能够建立其与天然组织的保真度。在这里,我们使用动物和体外模型组织的多维 NMR 光谱作为其各自分子结构的指纹,从而能够在原子长度尺度上比较完整的组织。为了从动物组织中获得光谱,我们开发了一种富含约 20%NMR 活性同位素碳-13 和氮-15 的重鼠。由此产生的光谱使我们能够改进正在发育的骨骼的体外模型,并探测其详细结构。鉴定出一种可能参与骨基质钙化的意想不到的分子聚(腺苷二磷酸核糖),说明了这种方法的分析能力。