title: Preparation of highly and generally enriched mammalian tissues for solid state NMR.
abstract: 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 (13)C 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.
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
authors.forename: Veronica Wai Ching
authors.initials: VW
authors.lastname: Wong
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
authors.forename: David G
authors.initials: DG
authors.lastname: Reid
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
authors.forename: Wing Ying
authors.initials: WY
authors.lastname: Chow
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
authors.forename: Rakesh
authors.initials: R
authors.lastname: Rajan
authors.affiliation: Central Biomedical Resources, School of Clinical Medicine, University of Cambridge, West Forvie Building, Forvie Site, Robinson Way, Cambridge, CB2 0SZ, UK.
authors.forename: Maggie
authors.initials: M
authors.lastname: Green
authors.affiliation: Department of Trauma and Orthopaedic Surgery, Addenbrooke's Hospital, University of Cambridge, Cambridge, CB2 0QQ, UK.
authors.forename: Roger A
authors.initials: RA
authors.lastname: Brooks
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK. mjd13@cam.ac.uk.
authors.forename: Melinda J
authors.initials: MJ
authors.lastname: Duer
mesh_terms: D000818:Animals; D051379:Mice; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D009928:Organ Specificity; D011506:Proteins / Q000737:chemistry*
chemical_list: D011506:Proteins
keywords: Bone collagen; Carbon-13; Fur keratin; In vivo; Labelling; Mouse