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Optimization of Metabolic Labeling with Alkyne-Containing Isoprenoid Probes

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Protein Lipidation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2009))

Abstract

Protein prenylation, found in eukaryotes, is a posttranslational modification in which one or two isoprenoid groups are added to the C terminus of selected proteins using either a farnesyl group or a geranylgeranyl group. Prenylation facilitates protein localization mainly to the plasma membrane where the prenylated proteins, including small GTPases, mediate signal transduction pathways. Changes in the level of prenylated proteins may serve a critical function in a variety of diseases. Metabolic labeling using modified isoprenoid probes followed by enrichment and proteomic analysis allows the identities and levels of prenylated proteins to be investigated. In this protocol, we illustrate how the conditions for metabolic labeling are optimized to maximize probe incorporation in HeLa cells through a combination of in-gel fluorescence and densitometric analysis.

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Acknowledgments

This work was supported in part by the National Institutes of Health (RF1AG056976 and GM084152) and by the National Science Foundation grant (CHE-1308655).

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Authors and Affiliations

  1. Department of Chemistry, University of Minnesota, Minneapolis, MN, USA

    Mina Ahmadi, Kiall Francis Suazo & Mark D. Distefano

Authors
  1. Mina Ahmadi
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  2. Kiall Francis Suazo
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  3. Mark D. Distefano
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Corresponding author

Correspondence to Mark D. Distefano .

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Editors and Affiliations

  1. Department of Molecular Medicine, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA

    Maurine E. Linder

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Ahmadi, M., Suazo, K.F., Distefano, M.D. (2019). Optimization of Metabolic Labeling with Alkyne-Containing Isoprenoid Probes. In: Linder, M. (eds) Protein Lipidation. Methods in Molecular Biology, vol 2009. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9532-5_3

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  • DOI: https://doi.org/10.1007/978-1-4939-9532-5_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9531-8

  • Online ISBN: 978-1-4939-9532-5

  • eBook Packages: Springer Protocols

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