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Investigation of Triple-Helix Collagen Hydroxylation by Solid-State NMR Spectroscopy

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Collagen

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1944))

Abstract

Solid-state nuclear magnetic resonance spectroscopy (ssNMR) is an emerging technique in structural methods of studying collagen proteins, capable of identifying features on an atomic length scale in tissues and protein samples without extensive extraction or purification. Hydroxylation is a key posttranslational modification of collagen that gives rise to distinctive signals in the ssNMR spectrum of collagen proteins. Here we outline the type of information that ssNMR can provide and describe the procedures involved in a ssNMR structural study, with particular focus on using dynamic nuclear polarization to enhance sensitivity for detecting hydroxylysine residues by ssNMR.

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Acknowledgments

The author thanks Dr. Jonathan Clark from the Babraham Institute for providing the lysine 13C-labelled mouse skin sample, Mr. Rakesh Rajan and Professor Melinda Duer from the University of Cambridge for providing the proline and glycine 13C, 15N-labelled fetal sheep osteoblast ECM sample, Mr. Robert Hayward and Professor Cathy Shanahan at King’s College London for providing the lysine 13C, 15N-labelled adult bovine vascular smooth muscle cell ECM sample, and Dr. Kelsey Collier at FMP Berlin for useful discussions and comments.

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Authors and Affiliations

  1. Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, Berlin, 13125, Germany

    Wing Ying Chow

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  1. Wing Ying Chow
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Correspondence to Wing Ying Chow .

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Editors and Affiliations

  1. Department of Biological Regulation, Weizmann Institute of Science, Rehovot, Israel

    Irit Sagi

  2. Department of Biological Regulation, Weizmann Institute of Science, Rehovot, Israel

    Nikolaos A. Afratis

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Chow, W.Y. (2019). Investigation of Triple-Helix Collagen Hydroxylation by Solid-State NMR Spectroscopy. In: Sagi, I., Afratis, N. (eds) Collagen. Methods in Molecular Biology, vol 1944. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9095-5_5

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  • DOI: https://doi.org/10.1007/978-1-4939-9095-5_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9094-8

  • Online ISBN: 978-1-4939-9095-5

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