Chimeric ZHHH neuroglobin acts as a cell membrane‐penetrating inducer of neurite outgrowth
Neuroglobin (Ngb) is a heme protein expressed in the vertebrate brain. We previously engineered a chimeric Ngb protein, in which module M1 of human Ngb is replaced by that of zebrafish Ngb, and showed that the chimeric ZHHH Ngb has a cell membrane‐penetrating activity similar to that of zebrafish Ng...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2017-09-01
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| Series: | FEBS Open Bio |
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| Online Access: | https://doi.org/10.1002/2211-5463.12271 |
| _version_ | 1828330573099171840 |
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| author | Nozomu Takahashi Wataru Onozuka Seiji Watanabe Keisuke Wakasugi |
| author_facet | Nozomu Takahashi Wataru Onozuka Seiji Watanabe Keisuke Wakasugi |
| author_sort | Nozomu Takahashi |
| collection | DOAJ |
| description | Neuroglobin (Ngb) is a heme protein expressed in the vertebrate brain. We previously engineered a chimeric Ngb protein, in which module M1 of human Ngb is replaced by that of zebrafish Ngb, and showed that the chimeric ZHHH Ngb has a cell membrane‐penetrating activity similar to that of zebrafish Ngb and also rescues cells from death caused by hypoxia/reoxygenation as does human Ngb. Recently, it was reported that overexpression of mammalian Ngb in neuronal cells induces neurite outgrowth. In this study, we performed neurite outgrowth assays of chimeric Ngb using rat pheochromocytoma PC12 cells. Addition of chimeric Ngb, but not human or zebrafish Ngb, exogenously to the cell medium induces neurite outgrowth. On the other hand, the K7A/K9Q chimeric Ngb double mutant, which cannot translocate into cells, did not induce neurite outgrowth, suggesting that the cell membrane‐penetrating activity of the chimeric Ngb is crucial for its neurite outgrowth‐promoting activity. We also prepared several site‐directed chimeric Ngb mutants and demonstrated that residues crucial for neurite outgrowth‐inducing activity of the chimeric Ngb are not exactly the same as those for its neuroprotective activity. |
| first_indexed | 2024-04-13T20:41:14Z |
| format | Article |
| id | doaj.art-f4f1879c072344a99f28e3025a52ae0b |
| institution | Directory Open Access Journal |
| issn | 2211-5463 |
| language | English |
| last_indexed | 2024-04-13T20:41:14Z |
| publishDate | 2017-09-01 |
| publisher | Wiley |
| record_format | Article |
| series | FEBS Open Bio |
| spelling | doaj.art-f4f1879c072344a99f28e3025a52ae0b2022-12-22T02:30:51ZengWileyFEBS Open Bio2211-54632017-09-01791338134910.1002/2211-5463.12271Chimeric ZHHH neuroglobin acts as a cell membrane‐penetrating inducer of neurite outgrowthNozomu Takahashi0Wataru Onozuka1Seiji Watanabe2Keisuke Wakasugi3Department of Life Sciences Graduate School of Arts and Sciences The University of Tokyo JapanDepartment of Life Sciences Graduate School of Arts and Sciences The University of Tokyo JapanDepartment of Life Sciences Graduate School of Arts and Sciences The University of Tokyo JapanDepartment of Life Sciences Graduate School of Arts and Sciences The University of Tokyo JapanNeuroglobin (Ngb) is a heme protein expressed in the vertebrate brain. We previously engineered a chimeric Ngb protein, in which module M1 of human Ngb is replaced by that of zebrafish Ngb, and showed that the chimeric ZHHH Ngb has a cell membrane‐penetrating activity similar to that of zebrafish Ngb and also rescues cells from death caused by hypoxia/reoxygenation as does human Ngb. Recently, it was reported that overexpression of mammalian Ngb in neuronal cells induces neurite outgrowth. In this study, we performed neurite outgrowth assays of chimeric Ngb using rat pheochromocytoma PC12 cells. Addition of chimeric Ngb, but not human or zebrafish Ngb, exogenously to the cell medium induces neurite outgrowth. On the other hand, the K7A/K9Q chimeric Ngb double mutant, which cannot translocate into cells, did not induce neurite outgrowth, suggesting that the cell membrane‐penetrating activity of the chimeric Ngb is crucial for its neurite outgrowth‐promoting activity. We also prepared several site‐directed chimeric Ngb mutants and demonstrated that residues crucial for neurite outgrowth‐inducing activity of the chimeric Ngb are not exactly the same as those for its neuroprotective activity.https://doi.org/10.1002/2211-5463.12271cell membrane‐penetrating proteinneurite outgrowthneuroglobinrat pheochromocytoma PC12 cell line |
| spellingShingle | Nozomu Takahashi Wataru Onozuka Seiji Watanabe Keisuke Wakasugi Chimeric ZHHH neuroglobin acts as a cell membrane‐penetrating inducer of neurite outgrowth FEBS Open Bio cell membrane‐penetrating protein neurite outgrowth neuroglobin rat pheochromocytoma PC12 cell line |
| title | Chimeric ZHHH neuroglobin acts as a cell membrane‐penetrating inducer of neurite outgrowth |
| title_full | Chimeric ZHHH neuroglobin acts as a cell membrane‐penetrating inducer of neurite outgrowth |
| title_fullStr | Chimeric ZHHH neuroglobin acts as a cell membrane‐penetrating inducer of neurite outgrowth |
| title_full_unstemmed | Chimeric ZHHH neuroglobin acts as a cell membrane‐penetrating inducer of neurite outgrowth |
| title_short | Chimeric ZHHH neuroglobin acts as a cell membrane‐penetrating inducer of neurite outgrowth |
| title_sort | chimeric zhhh neuroglobin acts as a cell membrane penetrating inducer of neurite outgrowth |
| topic | cell membrane‐penetrating protein neurite outgrowth neuroglobin rat pheochromocytoma PC12 cell line |
| url | https://doi.org/10.1002/2211-5463.12271 |
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