Unraveling the Mode of Action of a Neuron‐Specific Fluorescent Probe, NeuO: Intracellular Phosphorylation Through PAK6 Kinase

B Beomsue Kim (Neural Circuits Research Group Korea Brain Research Institute Daegu 41062 Republic of Korea) D Dongwan Ko (Neural Circuits Research Group Korea Brain Research Institute Daegu 41062 Republic of Korea) S Sang‐Bum Kim (New Drug Development Center Daegu–Gyeongbuk Medical Innovation Foundation (K‐MEDI Hub) Daegu 41061 Republic of Korea) S Songhui Kim (Neural Circuits Research Group Korea Brain Research Institute Daegu 41062 Republic of Korea) M Minji Kim N Na‐Kyeong Hong (Department of Chemistry Pohang University of Science and Technology (POSTECH) Pohang 37673 Republic of Korea) S Sun You Park (New Drug Development Center Daegu–Gyeongbuk Medical Innovation Foundation (K‐MEDI Hub) Daegu 41061 Republic of Korea) W Wonju Kim (Molecular Neurogenetics Unit, Center for Genomic Medicine, Massachusetts General Hospital) B Byoung‐Cheol Lee (Neurovascular Unit Research Group Korea Brain Research Institute Daegu 41062 Republic of Korea) Y Yunbeom Lee T Taebo Sim (Department of Biomedical Science, Graduate School of Medical Science, Brain Korea 21 Project Yonsei University College of Medicine Seoul 03722 Republic of Korea) Y Young‐Tae Chang (Department of Chemistry Pohang University of Science and Technology (POSTECH) Pohang 37673 Republic of Korea)

Abstract

Abstract NeuO is an intriguing molecular probe capable of selectively labeling live neurons within a heterogeneous population of brain cells. The precise mechanism responsible for this neuronal labeling has remained a longstanding enigma. We investigated four potential mechanisms underlying NeuO's fluorescence, including its interaction with neuronal proteins, transportation into neurons, export from glial cells, and structural alterations within neurons. Our study found that NeuO undergoes structural changes via phosphorylation by the PAK6 kinase, resulting in high fluorescence in neurons, likely due to fluorogenic activation and intracellular retention within neurons. This discovery improves our understanding of the molecular mechanism of NeuO in neurons and facilitates the development of new chemical compounds that can be selectively modified by kinases in living cells.

Article Details

Volume / Issue Vol. 64, Issue 37
Published September 08, 2025
ISSN 1433-7851
Publisher Wiley

Journal Info

Angewandte Chemie International Edition

Wiley

ISSN: 1433-7851 Physical Sciences

Authors (12)

B

Beomsue Kim

Neural Circuits Research Group Korea Brain Research Institute Daegu 41062 Republic of Korea

D

Dongwan Ko

Neural Circuits Research Group Korea Brain Research Institute Daegu 41062 Republic of Korea

S

Sang‐Bum Kim

New Drug Development Center Daegu–Gyeongbuk Medical Innovation Foundation (K‐MEDI Hub) Daegu 41061 Republic of Korea

S

Songhui Kim

Neural Circuits Research Group Korea Brain Research Institute Daegu 41062 Republic of Korea

M

Minji Kim

N

Na‐Kyeong Hong

Department of Chemistry Pohang University of Science and Technology (POSTECH) Pohang 37673 Republic of Korea

S

Sun You Park

New Drug Development Center Daegu–Gyeongbuk Medical Innovation Foundation (K‐MEDI Hub) Daegu 41061 Republic of Korea

W

Wonju Kim

Molecular Neurogenetics Unit, Center for Genomic Medicine, Massachusetts General Hospital

B

Byoung‐Cheol Lee

Neurovascular Unit Research Group Korea Brain Research Institute Daegu 41062 Republic of Korea

Y

Yunbeom Lee

T

Taebo Sim

Department of Biomedical Science, Graduate School of Medical Science, Brain Korea 21 Project Yonsei University College of Medicine Seoul 03722 Republic of Korea

Y

Young‐Tae Chang

Department of Chemistry Pohang University of Science and Technology (POSTECH) Pohang 37673 Republic of Korea