Using Steady-State Fluorescence Anisotropy to Study Protein Clustering

Ajeet Chaudhary; Kay Schneitz

doi:10.1007/978-1-0716-2132-5_16

Using Steady-State Fluorescence Anisotropy to Study Protein Clustering

Methods Mol Biol. 2022:2457:253-260. doi: 10.1007/978-1-0716-2132-5_16.

Authors

Ajeet Chaudhary^{1

2}, Kay Schneitz³

Affiliations

¹ Plant Developmental Biology, TUM School of Life Sciences, Technical University of Munich, Freising, Germany.
² Department of Plant Biology, Carnegie Institution for Science, Stanford, CA, USA.
³ Plant Developmental Biology, TUM School of Life Sciences, Technical University of Munich, Freising, Germany. kay.schneitz@tum.de.

PMID: 35349145
DOI: 10.1007/978-1-0716-2132-5_16

Abstract

Signaling pathways rely on the precise control of protein-protein interactions. Therefore, it is essential to be able to investigate such interactions with spatiotemporal resolution and in live cells. Here we describe a microscope-based fluorescence spectrometry technique to investigate homotypic interactions between GFP-labeled fusion proteins in a rapid and reproducible fashion using fluorescence anisotropy. This method is of great value for the study of protein complexes in live tissue with subcellular resolution.

Keywords: Arabidopsis; Cell wall; Confocal laser scanning microscopy; Fluorescence anisotropy; Methods; Microscopy; Plasmodesmata; Protein–protein interaction; Receptor kinase.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cluster Analysis
Fluorescence Polarization / methods
Proteins*
Spectrometry, Fluorescence

Substances

Proteins