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Detection of Protein Ubiquitination
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Detection of Protein Ubiquitination

Published on: August 19, 2009

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Assessment of ESCRT Protein CHMP5 Activity on Client Protein Ubiquitination by Immunoprecipitation and Western

Francheska Son1, Katharine Umphred-Wilson1, Jae-Hyuck Shim2

  • 1Department of Pathology, School of Medicine, Case Western Reserve University, Cleveland, OH, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 29, 2019
PubMed

Insights

Charged multivesicular body protein-5 (CHMP5) regulates protein ubiquitination in T lymphocytes. This study details a method to assess CHMP5

Area of Science:

  • Cell Biology
  • Immunology
  • Protein Biochemistry

Background:

  • Charged multivesicular body protein-5 (CHMP5) is an ESCRT component regulating membrane dynamics.
  • CHMP5 has emerging roles as a nonenzymatic regulator of protein ubiquitination and degradation.
  • Understanding CHMP5's function in immune cells is crucial for cellular regulation.

Purpose of the Study:

  • To describe a methodology for assessing CHMP5 activity.
  • To investigate CHMP5's role in regulating client protein ubiquitination.
  • To focus on CHMP5's function within T lymphocytes.

Main Methods:

  • Immunoprecipitation technique.
  • Western blot analysis.
  • Assessment of CHMP5 activity on protein ubiquitination in T lymphocytes.

Main Results:

  • The study presents a robust immunoprecipitation and western blot protocol.
  • This method allows for the evaluation of CHMP5's impact on client protein ubiquitination.
  • The methodology is specifically applied to T lymphocyte models.

Conclusions:

  • A novel methodology is established for studying CHMP5's non-ESCRT functions.
  • This technique facilitates the investigation of CHMP5's regulatory role in protein ubiquitination.
  • The findings provide a tool for deeper understanding of CHMP5 in immune cell signaling.
Keywords:
CHMP5DeubiquitinasePosttranslational modificationT lymphocytesUbiquitination

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