Coclustering of ErbB1 and ErbB2 revealed by FRET-sensitized acceptor bleaching.

Biophysical Journal
Agnes SzabóPeter Nagy

Abstract

Classical theory states that ligand binding induces the dimerization of ErbB proteins, leading to their activation. Although we and other investigators have shown the existence of preformed homoclusters of ErbB receptors and analyzed their composition, the stoichiometry of their heteroclusters has not been quantitatively described. Here, we report the development of the fluorescence resonance energy transfer (FRET)-sensitized acceptor bleaching (FSAB) technique to quantitate the ratio of ErbB1 and ErbB2 in their heteroclusters. In FSAB, photolabile acceptors within FRET distance from photostable donors are excited and photobleached by FRET, and the fraction of acceptors that are participating in FRET is determined. In quiescent SKBR-3 breast cancer cells, approximately 35% of ErbB1 and approximately 10% of ErbB2 have been found in heteroclusters. Epidermal growth factor (ligand of ErbB1) increased the fraction of ErbB2 heteroclustering with ErbB1, whereas the ratio of heteroclustered ErbB1 did not change significantly. The fractions of heteroclustered ErbB1 and ErbB2 were independent of their expression levels, indicating that the formation of these clusters is not driven by the law of mass action. In contrast, the FRET efficie...Continue Reading

References

Apr 29, 1999·Proceedings of the National Academy of Sciences of the United States of America·L N KlapperY Yarden
Nov 1, 2000·Cell·J Schlessinger
Mar 17, 2001·Nature Reviews. Molecular Cell Biology·Y Yarden, M X Sliwkowski
Dec 24, 2002·Biophysical Journal·Adam HoppeJoel A Swanson
Nov 5, 2003·Nature Biotechnology·Elizabeth A Jares-Erijman, Thomas M Jovin
Jul 3, 2004·European Biophysics Journal : EBJ·Andrew H A ClaytonEdouard C Nice
Mar 18, 2005·Journal of the American Chemical Society·Mike HeilemannPhilip Tinnefeld
Jan 25, 2006·Nature Methods·Kirsten BaciaPetra Schwille
Mar 3, 2006·The Journal of Physical Chemistry. a·Christian EggelingClaus A M Seidel
Jul 27, 2007·Journal of Cell Science·Shujie YangBridget S Wilson
Mar 8, 2008·The Journal of Physical Chemistry. B·Zeyu XiaoXiaohong Fang
May 23, 2008·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Corentin SprietLaurent Heliot
Jul 8, 2008·Angewandte Chemie·Jan VogelsangPhilip Tinnefeld
Sep 2, 2008·Experimental Cell Research·Ron Bose, Xuewu Zhang
Nov 29, 2008·Experimental Cell Research·Mark A Lemmon
Jan 27, 2009·Proceedings of the National Academy of Sciences of the United States of America·Michelle A DigmanEnrico Gratton
Aug 7, 2009·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Peter Nagy, János Szöllosi
Oct 6, 2009·Trends in Cell Biology·Diane S Lidke, Bridget S Wilson

❮ Previous
Next ❯

Citations

May 25, 2012·Nature Reviews. Cancer·Jessica B Casaletto, Andrea I McClatchey
Mar 31, 2015·International Journal of Molecular Sciences·Dilip ShresthaJános Szöllősi
Dec 17, 2015·The Journal of Chemical Physics·Jung Y Huang, Chien Y Lin
Nov 15, 2011·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Peter Nagy, János Szöllosi
Oct 22, 2016·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·János SzöllősiPéter Nagy
Jun 11, 2020·Methods and Applications in Fluorescence·Ágnes SzabóPeter Nagy

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.