FRAP beam-size analysis to measure palmitoylation-dependent membrane association dynamics and microdomain partitioning of Ras proteins

Yoav I. Henis, Barak Rotblat, Yoel Kloog

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

Abstract

Motions of membrane-associated proteins within and between membranes are essential for many cellular functions. We describe the application of fluorescence recovery after photobleaching (FRAP) beam-size analysis to investigate the role of palmitoylation in the membrane targeting and membrane association dynamics of H-Ras. The method described distinguishes between FRAP by lateral diffusion and by cytoplasmic exchange, and enables to obtain an estimate of the membrane affinity in live cells. These studies show distinct roles for the two palmitoylation sites (Cys181 and Cys184) on H-Ras, with different effects on membrane affinity and microlocalization.

Original languageEnglish
Pages (from-to)183-190
Number of pages8
JournalMethods
Volume40
Issue number2
DOIs
StatePublished - 1 Oct 2006
Externally publishedYes

Keywords

  • Beam-size analysis
  • Cholesterol
  • Exchange
  • FRAP
  • H-Ras
  • Lateral diffusion
  • Membrane association
  • Palmitoylation

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology (all)

Fingerprint

Dive into the research topics of 'FRAP beam-size analysis to measure palmitoylation-dependent membrane association dynamics and microdomain partitioning of Ras proteins'. Together they form a unique fingerprint.

Cite this