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Rapid stimulation of cellular Pi uptake by the inositol pyrophosphate InsP(8) induced by its photothermal release from lipid nanocarriers using a near infra-red light-emitting diode
Inositol pyrophosphates (PP-InsPs), including diphospho-myo-inositol pentakisphosphate (5-InsP(7)) and bis-diphospho-myo-inositol tetrakisphosphate (1,5-InsP(8)), are highly polar, membrane-impermeant signaling molecules that control many homeostatic responses to metabolic and bioenergetic imbalance...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Royal Society of Chemistry
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7891704/ https://www.ncbi.nlm.nih.gov/pubmed/33659052 http://dx.doi.org/10.1039/d0sc02144j |
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author | Wang, Zhenzhen Jork, Nikolaus Bittner, Tamara Wang, Huanchen Jessen, Henning J. Shears, Stephen B. |
author_facet | Wang, Zhenzhen Jork, Nikolaus Bittner, Tamara Wang, Huanchen Jessen, Henning J. Shears, Stephen B. |
author_sort | Wang, Zhenzhen |
collection | PubMed |
description | Inositol pyrophosphates (PP-InsPs), including diphospho-myo-inositol pentakisphosphate (5-InsP(7)) and bis-diphospho-myo-inositol tetrakisphosphate (1,5-InsP(8)), are highly polar, membrane-impermeant signaling molecules that control many homeostatic responses to metabolic and bioenergetic imbalance. To delineate their molecular activities, there is an increasing need for a toolbox of methodologies for real-time modulation of PP-InsP levels inside large populations of cultured cells. Here, we describe procedures to package PP-InsPs into thermosensitive phospholipid nanocapsules that are impregnated with a near infra-red photothermal dye; these liposomes are readily accumulated into cultured cells. The PP-InsPs remain trapped inside the liposomes until the cultures are illuminated with a near infra-red light-emitting diode (LED) which permeabilizes the liposomes to promote PP-InsP release. Additionally, so as to optimize these procedures, a novel stably fluorescent 5-InsP(7) analogue (i.e., 5-FAM-InsP(7)) was synthesized with the assistance of click-chemistry; the delivery and deposition of the analogue inside cells was monitored by flow cytometry and by confocal microscopy. We describe quantitatively-controlled PP-InsP release inside cells within 5 min of LED irradiation, without measurable effect upon cell integrity, using a collimated 22 mm beam that can irradiate up to 10(6) cultured cells. Finally, to interrogate the biological value of these procedures, we delivered 1,5-InsP(8) into HCT116 cells and showed it to dose-dependently stimulate the rate of [(33)P]-Pi uptake; these observations reveal a rheostatic range of concentrations over which 1,5-InsP(8) is biologically functional in Pi homeostasis. |
format | Online Article Text |
id | pubmed-7891704 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
spelling | pubmed-78917042021-03-02 Rapid stimulation of cellular Pi uptake by the inositol pyrophosphate InsP(8) induced by its photothermal release from lipid nanocarriers using a near infra-red light-emitting diode Wang, Zhenzhen Jork, Nikolaus Bittner, Tamara Wang, Huanchen Jessen, Henning J. Shears, Stephen B. Chem Sci Chemistry Inositol pyrophosphates (PP-InsPs), including diphospho-myo-inositol pentakisphosphate (5-InsP(7)) and bis-diphospho-myo-inositol tetrakisphosphate (1,5-InsP(8)), are highly polar, membrane-impermeant signaling molecules that control many homeostatic responses to metabolic and bioenergetic imbalance. To delineate their molecular activities, there is an increasing need for a toolbox of methodologies for real-time modulation of PP-InsP levels inside large populations of cultured cells. Here, we describe procedures to package PP-InsPs into thermosensitive phospholipid nanocapsules that are impregnated with a near infra-red photothermal dye; these liposomes are readily accumulated into cultured cells. The PP-InsPs remain trapped inside the liposomes until the cultures are illuminated with a near infra-red light-emitting diode (LED) which permeabilizes the liposomes to promote PP-InsP release. Additionally, so as to optimize these procedures, a novel stably fluorescent 5-InsP(7) analogue (i.e., 5-FAM-InsP(7)) was synthesized with the assistance of click-chemistry; the delivery and deposition of the analogue inside cells was monitored by flow cytometry and by confocal microscopy. We describe quantitatively-controlled PP-InsP release inside cells within 5 min of LED irradiation, without measurable effect upon cell integrity, using a collimated 22 mm beam that can irradiate up to 10(6) cultured cells. Finally, to interrogate the biological value of these procedures, we delivered 1,5-InsP(8) into HCT116 cells and showed it to dose-dependently stimulate the rate of [(33)P]-Pi uptake; these observations reveal a rheostatic range of concentrations over which 1,5-InsP(8) is biologically functional in Pi homeostasis. Royal Society of Chemistry 2020-09-08 /pmc/articles/PMC7891704/ /pubmed/33659052 http://dx.doi.org/10.1039/d0sc02144j Text en This journal is © The Royal Society of Chemistry 2020 http://creativecommons.org/licenses/by/3.0/ This article is freely available. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence (CC BY 3.0) |
spellingShingle | Chemistry Wang, Zhenzhen Jork, Nikolaus Bittner, Tamara Wang, Huanchen Jessen, Henning J. Shears, Stephen B. Rapid stimulation of cellular Pi uptake by the inositol pyrophosphate InsP(8) induced by its photothermal release from lipid nanocarriers using a near infra-red light-emitting diode |
title | Rapid stimulation of cellular Pi uptake by the inositol pyrophosphate InsP(8) induced by its photothermal release from lipid nanocarriers using a near infra-red light-emitting diode
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title_full | Rapid stimulation of cellular Pi uptake by the inositol pyrophosphate InsP(8) induced by its photothermal release from lipid nanocarriers using a near infra-red light-emitting diode
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title_fullStr | Rapid stimulation of cellular Pi uptake by the inositol pyrophosphate InsP(8) induced by its photothermal release from lipid nanocarriers using a near infra-red light-emitting diode
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title_full_unstemmed | Rapid stimulation of cellular Pi uptake by the inositol pyrophosphate InsP(8) induced by its photothermal release from lipid nanocarriers using a near infra-red light-emitting diode
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title_short | Rapid stimulation of cellular Pi uptake by the inositol pyrophosphate InsP(8) induced by its photothermal release from lipid nanocarriers using a near infra-red light-emitting diode
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title_sort | rapid stimulation of cellular pi uptake by the inositol pyrophosphate insp(8) induced by its photothermal release from lipid nanocarriers using a near infra-red light-emitting diode |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7891704/ https://www.ncbi.nlm.nih.gov/pubmed/33659052 http://dx.doi.org/10.1039/d0sc02144j |
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