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Hokkaido University Collection of Scholarly and Academic Papers >
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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
| 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)
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Submitter: 金城 政孝
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