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.
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.
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
Chika
C
Nakagawa
Shigenori
S
Nishimura
Kaori
K
Senda-Murata
Kenji
K
Sugimoto
D000818:Animals; D002477:Cells / Q000737:chemistry; D003584:Cytological Techniques / Q000379:methods*; D049452:Green Fluorescent Proteins; D057927:Hydrophobic and Hydrophilic Interactions; D008164:Luminescent Proteins; D008722:Methods; D020125:Mutation, Missense; D055503:Protein Multimerization; D011993:Recombinant Fusion Proteins; D028901:alpha Karyopherins / Q000235:genetics
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D028901:alpha Karyopherins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins