Nanoscale Organization of β2-Adrenergic Receptor-Venus Fusion Protein Domains on the Surface of Mammalian Cells

Download
  1. Get@NRC: Nanoscale Organization of β2-Adrenergic Receptor-Venus Fusion Protein Domains on the Surface of Mammalian Cells (Opens in a new window)
DOIResolve DOI: http://doi.org/10.1016/j.bbrc.2009.02.144
AuthorSearch for: ; Search for: ; Search for: ; Search for: ; Search for: ; Search for: ; Search for:
TypeArticle
Journal titleBiochemical and Biophysical Research Communications
Volume382
Issue1
Pages8590; # of pages: 6
SubjectAdrenergic receptor; GFP fusion; Inducible expression; Receptor signalling; Rafts
AbstractAdrenergic receptors are a key component of nanoscale multiprotein complexes that are responsible for controlling the beat rate in a mammalian heart. We demonstrate the ability of near-field scanning optical microscopy (NSOM) to visualize β2-adrenergic receptors (β2AR) fused to the GFP analogue Venus at the nanoscale on HEK293 cells. The expression of the β2AR-Venus fusion protein was tightly controlled using a tetracycline-induced promoter. Both the size and density of the observed nanoscale domains are dependent on the level of induction and thus the level of protein expression. At concentrations between 100 and 700 ng/ml of inducer doxycycline, the size of domains containing the β2AR-Venus fusion protein appears to remain roughly constant, but the number of domains per cell increase. At 700 ng/ml doxycycline the functional receptors are organized into domains with an average diameter of 150 nm with a density similar to that observed for the native protein on primary murine cells. By contrast, larger micron-sized domains of β2AR are observed in the membrane of the HEK293 cells that stably overexpress β2AR-GFP and β2AR-eYFP. We conclude that precise chemical control of gene expression is highly advantageous for the use β2AR-Venus fusion proteins as models for β2AR function. These observations are critical for designing future cell models and assays based on β2AR, since the receptor biology is consistent with a relatively low density of nanoscale receptor domains.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada (NRC-CNRC); NRC Steacie Institute for Molecular Sciences; NRC Institute for Biological Sciences
Peer reviewedYes
NRC publication
This is a non-NRC publication

"Non-NRC publications" are publications authored by NRC employees prior to their employment by NRC.

NPARC number16080421
Export citationExport as RIS
Report a correctionReport a correction
Record identifier8ad9c19b-ef74-4510-8395-f5d425fdf9f1
Record created2010-09-21
Record modified2016-05-09
Bookmark and share
  • Share this page with Facebook (Opens in a new window)
  • Share this page with Twitter (Opens in a new window)
  • Share this page with Google+ (Opens in a new window)
  • Share this page with Delicious (Opens in a new window)