Objectives: Prothymosin α (ProTα) was reported to inhibit the neuronal necrosis by facilitating the plasma membrane localization of endocytosed glucose transporter 1/4 through an activation of putative Gi-coupled receptor. The present study aims to identify a novel ProTα target, which may lead to an activation of Gi-coupled receptor. Methods: We used Gi-rich lipid rafts fraction of retinal cell line N18-RE-105 cells for affinity cross-linking. The biological confirmation that F0/F1 ATPase is a target protein complex was performed by cell-free experiments using ELISA-based binding assay, surface plasmon resonance assay and quartz crystal microbalance assay, and cell-based experiments to measure extracellular ATP level in the HUVECs culture. Results: From the cross-linking study and above-mentioned protein-protein interaction assays, ATP5A1 and ATP5B, F1 ATPase subunits were found to ProTα binding target proteins. In the culture of HUVEC cells, furthermore, ProTα increased the extracellular ATP levels in a reversible manner by anti-ATP5A1- and ATP5B-antibodies. Conclusion: The present study suggests that ProTα may activate ecto-F0/F1 ATPase and produced ATP. This study leads to next subjects whether produced ATP and its metabolites, ADP or adenosine may activate corresponding Gi-coupled receptors.
雑誌名
Expert Opinion on Biological Therapy
巻
18
号
Suppl 1
ページ
89 - 94
発行年
2018-07-31
出版者
Taylor and Francis Ltd
ISSN
14712598
EISSN
17447682
DOI
10.1080/14712598.2018.1454427
権利
c 2018 Informa UK Limited, trading as Taylor & Francis Group. This is an Accepted Manuscript of an article published by Taylor & Francis in Expert Opinion on Biological Therapy on 2018-07-31, available online: http://www.tandfonline.com/.10.1080/14712598.2018.1454427
著者版フラグ
author
引用
Expert Opinion on Biological Therapy, 18(suppl 1), pp.89-94; 2018
Ecto-F0/F1 ATPase as a novel candidate of prothymosin α receptor
EOBT18_89.pdf
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