Preparation of highly and generally enriched mammalian tissues for solid state NMR

详细信息    查看全文

• 作者：Veronica Wai Ching Wong ; David G. Reid ; Wing Ying Chow…
• 关键词：Carbon ; 13 ; In vivo ; Fur keratin ; Bone collagen ; Labelling ; Mouse
• 刊名：Journal of Biomolecular NMR
• 出版年：2015
• 出版时间：October 2015
• 年：2015
• 卷：63
• 期：2
• 页码：119-123
• 全文大小：1,069 KB
• 参考文献：Arnold AA et al (2013) Solid-state NMR structure determination of whole anchoring threads from the blue mussel Mytilus edulis. Biomacromolecules 14:132鈥?41CrossRef
  Berthold HK, Hachey DL, Reeds PJ, Thomas OP, Hoeksema S, Klein PD (1991) Uniformly C-13-labeled algal protein used to determine amino-acid essentiality in vivo. Proc Natl Acad Sci USA 88:8091鈥?095CrossRef ADS
  Chow WY et al (2014) NMR spectroscopy of native and in vitro tissues implicates polyADP ribose in biomineralization. Science 344:742鈥?46CrossRef ADS
  Hohwy M, Jakobsen HJ, Eden M, Levitt MH, Nielsen NC (1998) Broadband dipolar recoupling in the nuclear magnetic resonance of rotating solids: a compensated C7 pulse sequence. J Chem Phys 108:2686鈥?694CrossRef ADS
  McClatchy DB, Dong MQ, Wu CC, Venable JD, Yates JR (2007a) 15N metabolic labeling of mammalian tissue with slow protein turnover. J Proteome Res 6:2005鈥?010CrossRef
  McClatchy DB, Liao L, Park SK, Venable JD, Yates JR (2007b) Quantification of the synaptosomal proteome of the rat cerebellum during post-natal development. Genome Res 17:1378鈥?388CrossRef
  Nikel O, Laurencin D, McCallum SA, Gundberg CM, Vashishth D (2013) NMR investigation of the role of osteocalcin and osteopontin at the organic鈥搃norganic interface in bone. Langmuir 29:13873鈥?3882CrossRef
  Sarkar SK, Sullivan CE, Torchia DA (1983) Solid-state C-13 NMR study of collagen molecular-dynamics in hard and soft tissues. J Biol Chem 258:9762鈥?767
  Sarkar SK, Sullivan CE, Torchia DA (1985) Nanosecond fluctuations of the molecular backbone of collagen in hard and soft tissues鈥攁 C-13 nuclear magnetic resonance relaxation study. Biochemistry 24:2348鈥?354CrossRef
  Sarkar SK, Hiyama Y, Niu CH, Young PE, Gerig JT, Torchia DA (1987) Molecular-dynamics of collagen side-chains in hard and soft-tissues鈥攁 multinuclear magnetic-resonance study. Biochemistry 26:6793鈥?800CrossRef
  Saxena K, Dutta A, Klein-Seetharaman J, Schwalbe H (2012) Isotope labeling in insect cells. Methods Mol Biol 831:37鈥?4CrossRef
  Torchia DA, Hiyama Y, Sarkar SK, Sullivan CE, Young PE (1985) Multinuclear magnetic resonance studies of collagen molecular structure and dynamics. Biopolymers 24:65鈥?5CrossRef
  Verardi R, Traaseth NJ, Masterson LR, Vostrikov VV, Veglia G (2012) Isotope labeling for solution and solid-state NMR spectroscopy of membrane proteins. Adv Exp Med Biol 992:35鈥?2CrossRef
  Wagner G (2010) A topical issue: production and labeling of biological macromolecules for NMR investigations. J Biomol NMR 46:1鈥?CrossRef
  Wu CC, MacCoss MJ, Howell KE, Matthews DE, Yates JR 3rd (2004) Metabolic labeling of mammalian organisms with stable isotopes for quantitative proteomic analysis. Anal Chem 76:4951鈥?959CrossRef
  
• 作者单位：Veronica Wai Ching Wong (1)
  David G. Reid (1)
  Wing Ying Chow (1)
  Rakesh Rajan (1)
  Maggie Green (2)
  Roger A. Brooks (3)
  Melinda J. Duer (1)
  
  1. Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK
  2. Central Biomedical Resources, School of Clinical Medicine, University of Cambridge, West Forvie Building, Forvie Site, Robinson Way, Cambridge, CB2 0SZ, UK
  3. Department of Trauma and Orthopaedic Surgery, Addenbrooke鈥檚 Hospital, University of Cambridge, Cambridge, CB2 0QQ, UK
  
• 刊物类别：Physics and Astronomy
• 刊物主题：Physics
  Biophysics and Biomedical Physics
  Polymer Sciences
  Biochemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-5001

文摘

An appreciable level of isotope labelling is essential for future NMR structure elucidation of mammalian biomaterials, which are either poorly expressed, or unexpressable, using micro-organisms. We present a detailed protocol for high level 13C enrichment even in slow turnover murine biomaterials (fur keratin), using a customized diet supplemented with commercial labelled algal hydrolysate and formulated as a gel to minimize wastage, which female mice consumed during pregnancy and lactation. This procedure produced approximately eightfold higher fur keratin labelling in pups, exposed in utero and throughout life to label, than in adults exposed for the same period, showing both the effectiveness, and necessity, of this approach. Keywords Carbon-13 In vivo Fur keratin Bone collagen Labelling Mouse