Literature DB >> 24780222

Quantitative real-time kinetics of optogenetic proteins CRY2 and CIB1/N using single-molecule tools.

Yi Cui1, Samrat Roy Choudhury1, Joseph Irudayaraj2.   

Abstract

In this work we evaluate the interaction of two optogenetic protein variants (CIB1, CIBN) with their complementary protein CRY2 by single-molecule tools in cell-free extracts. After validating the blue light induced co-localization of CRY2 and CIB1/N by Förster resonance energy transfer (FRET) in live cells, a fluorescence correlation spectroscopy (FCS) based method was developed to quantitatively determine the in vitro association of the extracted proteins. Our experiments suggest that CIB1, in comparison with CIBN, possesses a better coupling efficiency with CRY2 due to its intact protein structure and lower diffusion rate within 300s detection window.
Copyright © 2014 Elsevier Inc. All rights reserved.

Entities:  

Keywords:  CIB1; CIBN; CRY2; FCS; FRET; Optogenetic protein

Mesh：

Substances：

Year:  2014        PMID： 24780222      PMCID： PMC4341968          DOI： 10.1016/j.ab.2014.04.023

Source DB:  PubMed          Journal:  Anal Biochem        ISSN： 0003-2697            Impact factor:   3.365


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