Function recovery after chemobleaching (FRAC): Evidence for activity silent membrane receptors on cell surface

Haiyan Sun, Sojin Shikano, Qiaojie Xiong, Min Li

Research output: Contribution to journalArticle

Abstract

Membrane proteins represent ≈30% of the proteome of both prokaryotes and eukaryotes. Unique to cell surface receptors is their biogenesis pathway, which involves vesicular trafficking from the endoplasmic reticulum through the Golgi apparatus and to the cell surface. Increasing evidence suggests specific regulation of biogenesis for different membrane receptors, hence affecting their surface expression. We report the development of a pulse-chase assay to monitor function recovery after chemobleaching (FRAC) to probe the transit time of the Kir2.1 K+ channel to reach the cell surface. Our results reveal that the channel activity is contributed by a small fraction of channel protein, providing evidence of activity-silent "sleeping" molecules on the cell surface. This method distinguishes molecular density from functional density, and the assay strategy is generally applicable to other membrane receptors. The ability of the reported method to access the biogenesis pathways in a high-throughput manner facilitates the identification and evaluation of molecules affecting receptor trafficking.

Original languageEnglish (US)
Pages (from-to)16964-16969
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume101
Issue number48
DOIs
StatePublished - Nov 30 2004

Keywords

  • Covalent modification
  • Fluorescence recovery after photobleaching
  • Ion channels
  • Signaling
  • Trafficking

ASJC Scopus subject areas

  • General

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