Monitoring RhoGDI extraction of lipid-modified Rho GTPases from membranes using click chemistry

Akiyuki Nishimura, Maurine E. Linder

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

The posttranslational lipid modification of Rho GTPases is important for their proper subcellular localization and signal transduction. Rho GTPases terminate in a CaaX motif, in which the cysteine residue is modified with either a farnesyl or geranylgeranyl isoprenoid. RhoGDI renders Rho GTPases soluble by masking their lipid moieties. We recently identified that the brain-specific splice variant of Cdc42 (bCdc42) containing a noncanonical CCaX motif harbors a dual prenyl–palmitoyl modification that prevents its binding to RhoGDI. This chapter describes a method to analyze RhoGDI extraction of Rho GTPases containing different lipid modifications from membranes using a liposome reconstitution assay and click chemistry.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages297-306
Number of pages10
DOIs
Publication statusPublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume2009
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Fingerprint

Click Chemistry
rho GTP-Binding Proteins
Lipids
Membranes
Terpenes
Post Translational Protein Processing
Membrane Lipids
Liposomes
Cysteine
Signal Transduction
Brain

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

Nishimura, A., & Linder, M. E. (2019). Monitoring RhoGDI extraction of lipid-modified Rho GTPases from membranes using click chemistry. In Methods in Molecular Biology (pp. 297-306). (Methods in Molecular Biology; Vol. 2009). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9532-5_22

Monitoring RhoGDI extraction of lipid-modified Rho GTPases from membranes using click chemistry. / Nishimura, Akiyuki; Linder, Maurine E.

Methods in Molecular Biology. Humana Press Inc., 2019. p. 297-306 (Methods in Molecular Biology; Vol. 2009).

Research output: Chapter in Book/Report/Conference proceedingChapter

Nishimura, A & Linder, ME 2019, Monitoring RhoGDI extraction of lipid-modified Rho GTPases from membranes using click chemistry. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 2009, Humana Press Inc., pp. 297-306. https://doi.org/10.1007/978-1-4939-9532-5_22
Nishimura A, Linder ME. Monitoring RhoGDI extraction of lipid-modified Rho GTPases from membranes using click chemistry. In Methods in Molecular Biology. Humana Press Inc. 2019. p. 297-306. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-9532-5_22
Nishimura, Akiyuki ; Linder, Maurine E. / Monitoring RhoGDI extraction of lipid-modified Rho GTPases from membranes using click chemistry. Methods in Molecular Biology. Humana Press Inc., 2019. pp. 297-306 (Methods in Molecular Biology).
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