Journal article
5-Nitro-2-(3-phenylpropylamino)benzoic Acid (NPPB) Stimulates Cellular ATP Release through Exocytosis of ATP-enriched Vesicles
The Journal of biological chemistry, Vol.284(49), pp.33894-33903
12/04/2009
DOI: 10.1074/jbc.M109.046193
PMCID: PMC2797160
PMID: 19808682
Abstract
Cells release ATP in response to physiologic stimuli. Extracellular ATP regulates a broad range of important cellular functions by activation of the purinergic receptors in the plasma membrane. The purpose of these studies was to assess the role of vesicular exocytosis in cellular ATP release. FM1-43 fluorescence was used to measure exocytosis and bioluminescence to measure ATP release in HTC rat hepatoma and Mz-Cha-1 human cholangiocarcinoma cells. Exposure to a Cl
−
channel inhibitor 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) (50–300 μ
m
) stimulated a 5–100-fold increase in extracellular ATP levels within minutes of the exposure. This rapid response was not a result of changes in cell viability or Cl
−
channel activity. NPPB also potently stimulated ATP release in HEK293 cells and HEK293 cells expressing a rat P2X7 receptor indicating that P2X7 receptors are not involved in stimulation of ATP release by NPPB. In all cells studied, NPPB rapidly stimulated vesicular exocytosis that persisted many minutes after the exposure. The kinetics of NPPB-evoked exocytosis and ATP release were similar. Furthermore, the magnitudes of NPPB-evoked exocytosis and ATP release were correlated (correlation coefficient 0.77), indicating that NPPB may stimulate exocytosis of a pool of ATP-enriched vesicles. These findings provide further support for the concept that vesicular exocytosis plays an important role in cellular ATP release and suggest that NPPB can be used as a biochemical tool to specifically stimulate ATP release through exocytic mechanisms.
Details
- Title: Subtitle
- 5-Nitro-2-(3-phenylpropylamino)benzoic Acid (NPPB) Stimulates Cellular ATP Release through Exocytosis of ATP-enriched Vesicles
- Creators
- Svjetlana Dolovcak - From the Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9151Shar L Waldrop - From the Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9151J. Gregory Fitz - From the Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9151Gordan Kilic - From the Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9151
- Resource Type
- Journal article
- Publication Details
- The Journal of biological chemistry, Vol.284(49), pp.33894-33903
- DOI
- 10.1074/jbc.M109.046193
- PMID
- 19808682
- PMCID
- PMC2797160
- NLM abbreviation
- J Biol Chem
- ISSN
- 0021-9258
- eISSN
- 1083-351X
- Publisher
- American Society for Biochemistry and Molecular Biology
- Grant note
- DK43278; DK46082 / National Institutes of Health
- Language
- English
- Date published
- 12/04/2009
- Academic Unit
- Immunology; Internal Medicine
- Record Identifier
- 9984094550102771
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