Real-Time In Vitro Fluorescence Anisotropy of the Cyanobacterial Circadian Clock

Joel Heisler; Archana Chavan; Yong-Gang Chang; Andy LiWang

doi:10.1007/978-1-0716-0381-9_1

Real-Time In Vitro Fluorescence Anisotropy of the Cyanobacterial Circadian Clock

Methods Mol Biol. 2021:2130:3-18. doi: 10.1007/978-1-0716-0381-9_1.

Authors

Joel Heisler^{1

2

3}, Archana Chavan³, Yong-Gang Chang^{1

3}, Andy LiWang^{4

5

6

7

8

9}

Affiliations

¹ Chemistry and Chemical Biology, University of California, Merced, CA, USA.
² Center for Cellular and Biomolecular Machines, University of California, Merced, CA, USA.
³ School of Natural Sciences, University of California, Merced, CA, USA.
⁴ Chemistry and Chemical Biology, University of California, Merced, CA, USA. aliwang@ucmerced.edu.
⁵ Center for Cellular and Biomolecular Machines, University of California, Merced, CA, USA. aliwang@ucmerced.edu.
⁶ School of Natural Sciences, University of California, Merced, CA, USA. aliwang@ucmerced.edu.
⁷ Center for Circadian Biology, University of California, La Jolla, CA, USA. aliwang@ucmerced.edu.
⁸ Quantitative and Systems Biology, University of California, Merced, CA, USA. aliwang@ucmerced.edu.
⁹ Health Sciences Research Institute, University of California, Merced, CA, USA. aliwang@ucmerced.edu.

PMID: 33284432
DOI: 10.1007/978-1-0716-0381-9_1

Abstract

Stochastic diffusion of a solution of fluorophores after photoselection reduces the polarization of emission, or fluorescence anisotropy. Because this randomization process is slower for larger molecules, fluorescence anisotropy is effective for measuring the kinetics of protein-binding events. Here, we describe how to use the technique to carry out real-time observations in vitro of the cyanobacterial circadian clock.

Keywords: Circadian clock; Cyanobacteria; Fluorescence; Phosphorylation; Protein.

Publication types

Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Circadian Clocks*
Cyanobacteria / genetics
Cyanobacteria / metabolism*
Fluorescence Polarization / methods

Grants and funding

R01 GM107521/GM/NIGMS NIH HHS/United States