A rapid and simple method of evaluating the dimeric tendency of fluorescent proteins in living cells using a truncated protein of importin α as fusion tag

Chika Nakagawa; Shigenori Nishimura; Kaori Senda-Murata; Kenji Sugimoto

doi:10.1271/bbb.110677

A rapid and simple method of evaluating the dimeric tendency of fluorescent proteins in living cells using a truncated protein of importin α as fusion tag

Biosci Biotechnol Biochem. 2012;76(2):388-90. doi: 10.1271/bbb.110677. Epub 2012 Feb 7.

Authors

Chika Nakagawa¹, Shigenori Nishimura, Kaori Senda-Murata, Kenji Sugimoto

Affiliation

¹ Laboratory of Molecular Biology and Cell Informatics, Division of Bioscience and Informatics, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Sakai, Osaka, Japan. cnakagawa@bioinfo.osakafu-u.ac.jp

PMID: 22313767
DOI: 10.1271/bbb.110677

Abstract

Enhanced green fluorescent protein (EGFP) and its yellow variant (Venus) are weakly dimeric under physiological conditions. We designed a simple method to evaluate the dimeric tendency of fluorescent proteins in living mammalian cells. A novel single mutation, A206L, interfering with the hydrophobic interactions of the dimer interface in Venus, contributed to its monomerization, and was as effective as the A206K mutation in this assay.

Publication types

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

MeSH terms

Animals
Cells / chemistry
Cytological Techniques / methods*
Green Fluorescent Proteins
Hydrophobic and Hydrophilic Interactions
Luminescent Proteins*
Methods
Mutation, Missense
Protein Multimerization*
Recombinant Fusion Proteins
alpha Karyopherins* / genetics

Substances

Luminescent Proteins
Recombinant Fusion Proteins
alpha Karyopherins
enhanced green fluorescent protein
Green Fluorescent Proteins