Bimolecular Fluorescence Complementation: Quantitative Analysis of In Cell Interaction of Nuclear Transporter Importin α with Cargo Proteins

Alexander Lee; Marie A Bogoyevitch; David A Jans

doi:10.1007/978-1-0716-2337-4_14

Bimolecular Fluorescence Complementation: Quantitative Analysis of In Cell Interaction of Nuclear Transporter Importin α with Cargo Proteins

Methods Mol Biol. 2022:2502:215-233. doi: 10.1007/978-1-0716-2337-4_14.

Authors

Alexander Lee^{1

2}, Marie A Bogoyevitch², David A Jans^{3

4}

Affiliations

¹ Nuclear Signalling Laboratory, Monash Biomedicine Discovery Institute, Monash University, Clayton, VIC, Australia.
² Department of Biochemistry and Molecular Biology, University of Melbourne, Parkville, VIC, Australia.
³ Nuclear Signalling Laboratory, Monash Biomedicine Discovery Institute, Monash University, Clayton, VIC, Australia. David.Jans@monash.edu.
⁴ Department of Biochemistry and Molecular Biology, University of Melbourne, Parkville, VIC, Australia. David.Jans@monash.edu.

PMID: 35412241
DOI: 10.1007/978-1-0716-2337-4_14

Abstract

Bimolecular fluorescence complementation utilizes the ability of two complementary nonfluorescent fragments to reconstitute and emit fluorescence when brought together through specific interaction of attached protein fragments of interest. It has been used in several different contexts to study protein-protein interaction. Here we apply the method for the first time to study interaction of the nuclear transporter importin α and its cargoes in a cellular context. By using image analysis to quantify the extent of nuclear complexation, it is possible to gain insight into the strength of interaction in cells.

Keywords: Bimolecular fluorescence complementation; CLSM; Importin α; Nuclear transport.

MeSH terms

Cell Communication* / physiology
Cell Nucleus / metabolism
Protein Binding
Proteins* / metabolism
Spectrometry, Fluorescence* / methods
alpha Karyopherins* / metabolism

Substances

Proteins
alpha Karyopherins