Preparation of highly and generally enriched mammalian tissues for solid state NMR.
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Authors
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.
Description
Keywords
Bone collagen, Carbon-13, Fur keratin, In vivo, Labelling, Mouse, Animals, Mice, Nuclear Magnetic Resonance, Biomolecular, Organ Specificity, Proteins
Journal Title
J Biomol NMR
Conference Name
Journal ISSN
0925-2738
1573-5001
1573-5001
Volume Title
63
Publisher
Springer Science and Business Media LLC
Publisher DOI
Sponsorship
Biotechnology and Biological Sciences Research Council (BB/G021392/1)
The authors would like to acknowledge funding from the Biotechnology and Biological Sciences Research Council for DGR and RR; Engineering and Physical Sciences Research Council for WYC and VWCW; National Institute of Health Research for RAB.