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Determination of dissociation constant of the NF kappa B p50/p65 heterodimer using fluorescence cross-correlation spectroscopy in the living cell

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Title: Determination of dissociation constant of the NF kappa B p50/p65 heterodimer using fluorescence cross-correlation spectroscopy in the living cell
Authors: Tiwari, Manisha Browse this author
Mikuni, Shintaro Browse this author
Muto, Hideki Browse this author
Kinjo, Masataka Browse this author →KAKEN DB
Keywords: NFkB
p50/p65
Dissociation constant
Heterodimer
Fluorescence measurement
FCS/FCCS
Live cell imaging
Issue Date: 5-Jul-2013
Publisher: Elsevier
Journal Title: Biochemical and Biophysical Research Communications
Volume: 436
Issue: 3
Start Page: 430
End Page: 435
Publisher DOI: 10.1016/j.bbrc.2013.05.121
PMID: 23751347
Abstract: Two-laser-beam fluorescence cross-correlation spectroscopy (FCCS) is promising technique that provides quantitative information about the interactions of biomolecules. The p50/p65 heterodimer is the most abundant and well understood of the NF kappa B dimers in most cells. However, the quantitative value of affinity, namely the K-d, for the heterodimer in living cells is not known yet. To quantify the heterodimerization of the IPT domain of p50/p65 in the living cell, we used two-laser-beam FCCS. The K-d values of mCherry(2)- and EGFP-fused p50 and p65 were determined to be 0.46 mu M in the cytoplasm and 1.06 mu M in the nucleus of the living cell. These results suggest the different binding affinities of the p50/p65 heterodimer in the cytoplasm and nucleus of the living cell and different complex formation in each region. (C) 2013 Elsevier Inc. All rights reserved.
Type: article (author version)
URI: http://hdl.handle.net/2115/53136
Appears in Collections:生命科学院・先端生命科学研究院 (Graduate School of Life Science / Faculty of Advanced Life Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 金城 政孝

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