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

Biochemistry & Molecular Biology Notes

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

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Keywords: fluorescent protein (FP), Venus, weak dimer, importin alpha (importin α), nuclear-pore complex (NPC)

JOURNAL FREE ACCESS

2012 Volume 76 Issue 2 Pages 388-390

DOI https://doi.org/10.1271/bbb.110677

View "Advance Publication" version (February 7, 2012).

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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.

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