A genetically encoded toolkit for tracking live-cell histidine dynamics in space and time.
Sci Rep
; 7: 43479, 2017 03 02.
Article
em En
| MEDLINE
| ID: mdl-28252043
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.
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Base de dados:
MEDLINE
Assunto principal:
Proteínas de Bactérias
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Técnicas Biossensoriais
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Membrana Celular
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Proteínas Periplásmicas de Ligação
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Membranas Mitocondriais
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Histidina
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Proteínas Luminescentes
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article