Specific Fluorescent Probe for Protein Histidine Phosphatase Activity
Protein histidine phosphorylation plays a vital role in cell signaling and metabolic processes, and phosphohistidine (pHis) phosphatases such as protein histidine phosphatase 1 (PHPT1) and LHPP have been linked to cancer and diabetes, making them novel drug targets and biomarkers. Unlike the case for other classes of phosphatases, further studies of PHPT1 and other pHis phosphatases have been hampered by the lack of specific activity assays in complex biological mixtures. Previous methods relying on radiolabeling are hazardous and technically laborious, and small-molecule phosphatase probes are not selective toward pHis phosphatases. To address these issues, we herein report a fluorescent probe based on chelation-enhanced fluorescence (CHEF) to continuously measure the pHis phosphatase activity of PHPT1. Our probe exhibited excellent sensitivity and specificity toward PHPT1, enabling the first specific measurement of PHPT1 activity in cell lysates. Using this probe, we also obtained more physiologically relevant kinetic parameters of PHPT1, overcoming the limitations of previously used methods.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2019 |
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| Erschienen: |
2019 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:4 |
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| Enthalten in: |
ACS sensors - 4(2019), 4 vom: 26. Apr., Seite 1055-1062 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Choi, Yigun [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 24.04.2020 Date Revised 24.04.2020 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1021/acssensors.9b00242 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM295350202 |
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| 245 | 1 | 0 | |a Specific Fluorescent Probe for Protein Histidine Phosphatase Activity |
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| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Protein histidine phosphorylation plays a vital role in cell signaling and metabolic processes, and phosphohistidine (pHis) phosphatases such as protein histidine phosphatase 1 (PHPT1) and LHPP have been linked to cancer and diabetes, making them novel drug targets and biomarkers. Unlike the case for other classes of phosphatases, further studies of PHPT1 and other pHis phosphatases have been hampered by the lack of specific activity assays in complex biological mixtures. Previous methods relying on radiolabeling are hazardous and technically laborious, and small-molecule phosphatase probes are not selective toward pHis phosphatases. To address these issues, we herein report a fluorescent probe based on chelation-enhanced fluorescence (CHEF) to continuously measure the pHis phosphatase activity of PHPT1. Our probe exhibited excellent sensitivity and specificity toward PHPT1, enabling the first specific measurement of PHPT1 activity in cell lysates. Using this probe, we also obtained more physiologically relevant kinetic parameters of PHPT1, overcoming the limitations of previously used methods | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a PHPT1 | |
| 650 | 4 | |a chelation-enhanced fluorescence | |
| 650 | 4 | |a enzyme kinetics | |
| 650 | 4 | |a fluorescent probe | |
| 650 | 4 | |a phosphohistidine phosphatase | |
| 650 | 7 | |a Fluorescent Dyes |2 NLM | |
| 650 | 7 | |a Quinolines |2 NLM | |
| 650 | 7 | |a Sulfonamides |2 NLM | |
| 650 | 7 | |a PHPT1 protein, human |2 NLM | |
| 650 | 7 | |a EC 3.1.3.- |2 NLM | |
| 650 | 7 | |a Phosphoric Monoester Hydrolases |2 NLM | |
| 650 | 7 | |a EC 3.1.3.2 |2 NLM | |
| 650 | 7 | |a Inorganic Pyrophosphatase |2 NLM | |
| 650 | 7 | |a EC 3.6.1.1 |2 NLM | |
| 650 | 7 | |a phospholysine phosphohistidine inorganic pyrophosphate phosphatase, human |2 NLM | |
| 650 | 7 | |a EC 3.6.1.1 |2 NLM | |
| 700 | 1 | |a Shin, Son Hye |e verfasserin |4 aut | |
| 700 | 1 | |a Jung, Hoyoung |e verfasserin |4 aut | |
| 700 | 1 | |a Kwon, Ohyeon |e verfasserin |4 aut | |
| 700 | 1 | |a Seo, Jeong Kon |e verfasserin |4 aut | |
| 700 | 1 | |a Kee, Jung-Min |e verfasserin |4 aut | |
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