Folding Latency of Fluorescent Proteins Affects the Mitochondrial Localization of Fusion Proteins.

Abstract:

:The discovery of fluorescent proteins (FPs) has revolutionized cell biology. The fusion of targeting sequences to FPs enables the investigation of cellular organelles and their dynamics; however, occasionally, such fluorescent fusion proteins (FFPs) exhibit behavior different from that of the native proteins. Here, we constructed a color pallet comprising different organelle markers and found that FFPs targeted to the mitochondria were mislocalized when fused to certain types of FPs. Such FPs included several variants of Aequorea victoria green FP (avGFP) and a monomeric variant of the red FP. Because the FFPs that are mislocalized include FPs with faster maturing or folding mutations, the increase in the maturation rate is likely to prevent their expected localization. Indeed, when we reintroduced amino acid substitutions so that the FP sequences were equivalent to that of wild-type avGFP, FFP localization to the mitochondria was significantly enhanced. Moreover, similar amino acid substitutions improved the localization of mitochondria-targeted pHluorin, which is a pH-sensitive variant of GFP, and its capability to monitor pH changes in the mitochondrial matrix. Our findings demonstrate the importance of selecting FPs that maximize FFP function.Key words: fluorescent protein, organelle, fusion protein, mitochondria.

journal_name

Cell Struct Funct

authors

Kashiwagi S,Fujioka Y,Satoh AO,Yoshida A,Fujioka M,Nepal P,Tsuzuki A,Aoki O,Paudel S,Sasajima H,Ohba Y

doi

10.1247/csf.19028

subject

Has Abstract

pub_date

2019-12-26 00:00:00

pages

183-194

issue

2

eissn

0386-7196

issn

1347-3700

journal_volume

44

pub_type

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