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

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.

SUBMITTER: Hu H

PROVIDER: S-EPMC5333150 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature

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

Hu Hanyang H Gu Yanfang Y Xu Lei L Zou Yejun Y Wang Aoxue A Tao Rongkun R Chen Xianjun X Zhao Yuzheng Y Yang Yi Y

Scientific reports 20170302

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 his ...[more]

PMID: 28252043

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