Open Access System for Information Sharing

Login Library

 

Article
Cited 0 time in webofscience Cited 0 time in scopus
Metadata Downloads

Pairwise detection of site-specific receptor phosphorylations using

Title
Pairwise detection of site-specific receptor phosphorylations using
Authors
Kim, KLKim, DLee, SKim, SJNoh, JEKim, JHChae, YCLee, JBRyu, SH
POSTECH Authors
Kim, JHLee, JBRyu, SH
Date Issued
Mar-2016
Publisher
Nature Publishing Group
Abstract
Post-translational modifications (PTMs) of receptor tyrosine kinases (RTKs) at the plasma membrane (PM) determine the signal transduction efficacy alone and in combination. However, current approaches to identify PTMs provide ensemble results, inherently overlooking combinatorial PTMs in a single polypeptide molecule. Here, we describe a single-molecule blotting (SiMBlot) assay that combines biotinylation of cell surface receptors with single-molecule fluorescence microscopy. This method enables quantitative measurement of the phosphorylation status of individual membrane receptor molecules and colocalization analysis of multiple immunofluorescence signals to directly visualize pairwise site-specific phosphorylation patterns at the single-molecule level. Strikingly, application of SiMBlot to study ligand-dependent epidermal growth factor receptor (EGFR) phosphorylation, which is widely thought to be multi-phosphorylated, reveals that EGFR on cell membranes is hardly multi-phosphorylated, unlike in vitro autophosphorylated EGFR. Therefore, we expect SiMBlot to aid understanding of vast combinatorial PTM patterns, which are concealed in ensemble methods, and to broaden knowledge of RTK signaling.
URI
http://oasis.postech.ac.kr/handle/2014.oak/29995
DOI
10.1038/NCOMMS11107
ISSN
2041-1723
Article Type
Article
Citation
NATURE COMMUNICATIONS, vol. 7, page. 11107, 2016-03
Files in This Item:

qr_code

  • mendeley

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

Related Researcher

Researcher

 LEE, JONG BONG
Dept of Physics
Read more

Altmetric

Views & Downloads

Browse