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Bioconjugation pp 149–165Cite as

Protein Labeling and Bioconjugation Using N-Myristoyltransferase

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2033))

Abstract

Methods that allow for labeling of proteins cotranslationally within protein expression systems have had wide-ranging applications in health, engineering, and medicine. Bioorthogonal chemistries that allow for conjugation of proteins or biomolecules of interest to substrates (fluorophores, gold nanoparticles, polymers, etc.) in living cells without prior enrichment or purification have likewise enabled advances in technology to study and engineer cellular and biomolecular systems. At the intersection of these, chemoenzymatic labeling of proteins at specific sites of interest and their subsequent selective bioconjugation to substrates without prior purification has dramatically streamlined workflows that allow proteins to reside in the native expression volumes as long as possible prior to conjugation, be readily isolated upon conjugation, and remain functionally active after conjugation. Here we present methods and protocols to express and label proteins of interest at the N-terminus with azide derivatives of myristic acid, a small, soluble, 14-carbon fatty acid, and conjugate the labeled protein to fluorophores and gold nanoparticle substrates. These methods can be extended to label proteins with other myristoyl derivatives and to conjugation to other solid or polymeric substrates of interest.

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Acknowledgments

The authors wish to thank Dr. Katherine N. Clayton who has been instrumental in developing the protocols described here.

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Authors and Affiliations

  1. Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, USA

    Karin F. K. Ejendal, Julia G. Fraseur & Tamara L. Kinzer-Ursem

Authors
  1. Karin F. K. Ejendal
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  2. Julia G. Fraseur
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  3. Tamara L. Kinzer-Ursem
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Corresponding author

Correspondence to Tamara L. Kinzer-Ursem .

Editor information

Editors and Affiliations

  1. Cellular and Molecular Immunology Laboratory, Vrije Universiteit Brussel, Brussels, Belgium

    Sam Massa

  2. In Vivo Cellular and Molecular Imaging Laboratory (ICMI), Vrije Universiteit Brussel, Brussels, Belgium

    Nick Devoogdt

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Ejendal, K.F.K., Fraseur, J.G., Kinzer-Ursem, T.L. (2019). Protein Labeling and Bioconjugation Using N-Myristoyltransferase. In: Massa, S., Devoogdt, N. (eds) Bioconjugation. Methods in Molecular Biology, vol 2033. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9654-4_11

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  • DOI: https://doi.org/10.1007/978-1-4939-9654-4_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9653-7

  • Online ISBN: 978-1-4939-9654-4

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