A genetically encoded toolkit for tracking live-cell histidine dynamics in space and time

Hanyang Hu; Yanfang Gu; Lei Xu; Yejun Zou; Aoxue Wang; Rongkun Tao; Xianjun Chen; Yuzheng Zhao; Yi Yang

doi:10.1038/srep43479

A genetically encoded toolkit for tracking live-cell histidine dynamics in space and time

Sci Rep. 2017 Mar 2:7:43479. doi: 10.1038/srep43479.

Authors

Hanyang Hu^{1

2

3}, Yanfang Gu^{1

2

3}, Lei Xu^{1

2}, Yejun Zou^{1

2

3}, Aoxue Wang^{1

2

3}, Rongkun Tao^{1

2

3}, Xianjun Chen^{1

2

3}, Yuzheng Zhao^{1

2

3}, Yi Yang^{1

2}

Affiliations

¹ Synthetic Biology and Biotechnology Laboratory, State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, East China University of Science and Technology, 130 Mei Long Road, Shanghai 200237, China.
² Optogenetics &Molecular Imaging Interdisciplinary Research Center, CAS Center for Excellence in Brain Science, East China University of Science and Technology, 130 Mei Long Road, Shanghai 200237, China.
³ Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, 130 Mei Long Road, Shanghai 200237, China.

Abstract

High-resolution spatiotemporal imaging of histidine in single living mammalian cells faces technical challenges. Here, we developed a series of ratiometric, highly responsive, and single fluorescent protein-based histidine sensors of wide dynamic range. We used these sensors to quantify subcellular free-histidine concentrations in glucose-deprived cells and glucose-fed cells. Results showed that cytosolic free-histidine concentration was higher and more sensitive to the environment than free histidine in the mitochondria. Moreover, histidine was readily transported across the plasma membrane and mitochondrial inner membrane, which had almost similar transport rates and transport constants, and histidine transport was not influenced by cellular metabolic state. These sensors are potential tools for tracking histidine dynamics inside subcellular organelles, and they will open an avenue to explore complex histidine signaling.

Publication types

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

MeSH terms

Bacterial Proteins / chemistry*
Bacterial Proteins / genetics
Bacterial Proteins / metabolism
Binding Sites
Biological Transport
Biosensing Techniques*
Cell Membrane / drug effects
Cell Membrane / metabolism*
Cell Tracking
Cloning, Molecular
Cytosol / drug effects
Cytosol / metabolism
Escherichia coli / genetics
Escherichia coli / metabolism
Fluorescence Resonance Energy Transfer
Gene Expression
Glucose / metabolism
Glucose / pharmacology
HeLa Cells
Histidine / analysis*
Histidine / metabolism
Humans
Luminescent Proteins / chemistry*
Luminescent Proteins / genetics
Luminescent Proteins / metabolism
Mitochondria / drug effects
Mitochondria / metabolism
Mitochondrial Membranes / drug effects
Mitochondrial Membranes / metabolism*
Periplasmic Binding Proteins / chemistry*
Periplasmic Binding Proteins / genetics
Periplasmic Binding Proteins / metabolism
Protein Binding
Protein Conformation, alpha-Helical
Protein Conformation, beta-Strand
Protein Interaction Domains and Motifs
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Recombinant Proteins / metabolism
Signal Transduction

Substances

Bacterial Proteins
Luminescent Proteins
Periplasmic Binding Proteins
Recombinant Proteins
histidine-binding protein
yellow fluorescent protein, Bacteria
Histidine
Glucose