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


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Authors

Wong, Veronica Wai Ching 
Reid, David G 
Chow, Wing Ying 
Rajan, Rakesh 
Green, Maggie 

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

Volume Title

63

Publisher

Springer Science and Business Media LLC
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.