Cellular phosphatidic acid sensor, α-synuclein N-terminal domain, detects endogenous phosphatidic acid in macrophagic phagosomes and neuronal growth cones
Phosphatidic acid (PA) is the simplest phospholipid and is involved in the regulation of various cellular events. Recently, we developed a new PA sensor, the N-terminal region of α-synuclein (α-Syn-N). However, whether α-Syn-N can sense physiologically produced, endogenous PA remains unclear. We fir...
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| Format: | Article |
| Language: | English |
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Elsevier
2020-07-01
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| Series: | Biochemistry and Biophysics Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405580820300789 |
| _version_ | 1818180726049210368 |
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| author | Haruka Yamada Fumi Hoshino Qiang Lu Fumio Sakane |
| author_facet | Haruka Yamada Fumi Hoshino Qiang Lu Fumio Sakane |
| author_sort | Haruka Yamada |
| collection | DOAJ |
| description | Phosphatidic acid (PA) is the simplest phospholipid and is involved in the regulation of various cellular events. Recently, we developed a new PA sensor, the N-terminal region of α-synuclein (α-Syn-N). However, whether α-Syn-N can sense physiologically produced, endogenous PA remains unclear. We first established an inactive PA sensor (α-Syn-N-KQ) as a negative control by replacing all eleven lysine residues with glutamine residues. Using confocal microscopy, we next verified that α-Syn-N, but not α-Syn-N-KQ, detected PA in macrophagic phagosomes in which PA is known to be enriched, further indicating that α-Syn-N can be used as a reliable PA sensor in cells. Finally, because PA generated during neuronal differentiation is critical for neurite outgrowth, we investigated the subcellular distribution of PA using α-Syn-N. We found that α-Syn-N, but not α-Syn-N-KQ, accumulated at the peripheral regions (close to the plasma membrane) of neuronal growth cones. Experiments using a phospholipase D (PLD) inhibitor strongly suggested that PA in the peripheral regions of the growth cone was primarily produced by PLD. Our findings provide a reliable sensor of endogenous PA and novel insights into the distribution of PA during neuronal differentiation. |
| first_indexed | 2024-12-11T21:24:21Z |
| format | Article |
| id | doaj.art-0ec1b1ac2429461eabeb32b2e0e5ec94 |
| institution | Directory Open Access Journal |
| issn | 2405-5808 |
| language | English |
| last_indexed | 2024-12-11T21:24:21Z |
| publishDate | 2020-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biochemistry and Biophysics Reports |
| spelling | doaj.art-0ec1b1ac2429461eabeb32b2e0e5ec942022-12-22T00:50:22ZengElsevierBiochemistry and Biophysics Reports2405-58082020-07-0122Cellular phosphatidic acid sensor, α-synuclein N-terminal domain, detects endogenous phosphatidic acid in macrophagic phagosomes and neuronal growth conesHaruka Yamada0Fumi Hoshino1Qiang Lu2Fumio Sakane3Department of Chemistry, Graduate School of Science, Chiba University, Chiba, 263-8522, JapanDepartment of Chemistry, Graduate School of Science, Chiba University, Chiba, 263-8522, JapanDepartment of Chemistry, Graduate School of Science, Chiba University, Chiba, 263-8522, JapanCorresponding author.Department of Chemistry, Graduate School of Science, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba, 263-8522, Japan.; Department of Chemistry, Graduate School of Science, Chiba University, Chiba, 263-8522, JapanPhosphatidic acid (PA) is the simplest phospholipid and is involved in the regulation of various cellular events. Recently, we developed a new PA sensor, the N-terminal region of α-synuclein (α-Syn-N). However, whether α-Syn-N can sense physiologically produced, endogenous PA remains unclear. We first established an inactive PA sensor (α-Syn-N-KQ) as a negative control by replacing all eleven lysine residues with glutamine residues. Using confocal microscopy, we next verified that α-Syn-N, but not α-Syn-N-KQ, detected PA in macrophagic phagosomes in which PA is known to be enriched, further indicating that α-Syn-N can be used as a reliable PA sensor in cells. Finally, because PA generated during neuronal differentiation is critical for neurite outgrowth, we investigated the subcellular distribution of PA using α-Syn-N. We found that α-Syn-N, but not α-Syn-N-KQ, accumulated at the peripheral regions (close to the plasma membrane) of neuronal growth cones. Experiments using a phospholipase D (PLD) inhibitor strongly suggested that PA in the peripheral regions of the growth cone was primarily produced by PLD. Our findings provide a reliable sensor of endogenous PA and novel insights into the distribution of PA during neuronal differentiation.http://www.sciencedirect.com/science/article/pii/S2405580820300789Phosphatidic acidLipid sensorα-SynucleinPhagosomeGrowth conePhospholipase D |
| spellingShingle | Haruka Yamada Fumi Hoshino Qiang Lu Fumio Sakane Cellular phosphatidic acid sensor, α-synuclein N-terminal domain, detects endogenous phosphatidic acid in macrophagic phagosomes and neuronal growth cones Biochemistry and Biophysics Reports Phosphatidic acid Lipid sensor α-Synuclein Phagosome Growth cone Phospholipase D |
| title | Cellular phosphatidic acid sensor, α-synuclein N-terminal domain, detects endogenous phosphatidic acid in macrophagic phagosomes and neuronal growth cones |
| title_full | Cellular phosphatidic acid sensor, α-synuclein N-terminal domain, detects endogenous phosphatidic acid in macrophagic phagosomes and neuronal growth cones |
| title_fullStr | Cellular phosphatidic acid sensor, α-synuclein N-terminal domain, detects endogenous phosphatidic acid in macrophagic phagosomes and neuronal growth cones |
| title_full_unstemmed | Cellular phosphatidic acid sensor, α-synuclein N-terminal domain, detects endogenous phosphatidic acid in macrophagic phagosomes and neuronal growth cones |
| title_short | Cellular phosphatidic acid sensor, α-synuclein N-terminal domain, detects endogenous phosphatidic acid in macrophagic phagosomes and neuronal growth cones |
| title_sort | cellular phosphatidic acid sensor α synuclein n terminal domain detects endogenous phosphatidic acid in macrophagic phagosomes and neuronal growth cones |
| topic | Phosphatidic acid Lipid sensor α-Synuclein Phagosome Growth cone Phospholipase D |
| url | http://www.sciencedirect.com/science/article/pii/S2405580820300789 |
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