This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Arakaki, N. Articles by Higuti, T. Search for Related Content PubMed PubMed Citation Articles by Arakaki, N. Articles by Higuti, T.

---

Angiogenesis, Metastasis, and the Cellular Microenvironment

Possible Role of Cell Surface H⁺-ATP Synthase in the Extracellular ATP Synthesis and Proliferation of Human Umbilical Vein Endothelial Cells¹

¹ Faculty of Pharmaceutical Sciences, University of Tokushima, Tokushima, Japan and
² Daiichi Suntory Biomedical Research Co., Ltd., Osaka, Japan

Requests for reprints: Naokatu Arakaki, Faculty of Pharmaceutical Sciences, University of Tokushima, Shomachi, Tokushima 770-8505, Japan. Phone: 81-88-633-7255; Fax: 81-88-633-9550. E-mail: arakaki{at}ph.tokushima-u.ac.jp

Extracellular ATP synthesis on human umbilical vein endothelial cells (HUVECs) was examined, and it was found that HUVECs possess high ATP synthesis activity on the cell surface. Extracellular ATP generation was detected within 5 s after addition of ADP and inorganic phosphate and reached a maximal level at 15 s. This type of ATP synthesis was almost completely inhibited by mitochondrial H⁺-ATP synthase inhibitors (e.g., efrapeptins, resveratrol, and piceatannol), which target the F₁ catalytic domain. Oligomycin and carbonyl cyanide m-chlorophenylhydrazone, but not potassium cyanide, also inhibited extracellular ATP synthesis on HUVECs, suggesting that cell surface ATP synthase employs the transmembrane electrochemical potential difference of protons to synthesize ATP as well as mitochondrial H⁺-ATP synthase. The F₁-targeting H⁺-ATP synthase inhibitors markedly inhibited the proliferation of HUVECs, but intracellular ATP levels in HUVECs treated with these inhibitors were only slightly affected, as shown by comparison with the control cells. Interestingly, piceatannol inhibited only partially the activation of Syk (a nonreceptor tyrosine kinase), which has been shown to play a role in a number of endothelial cell functions, including cell growth and migration. These findings suggest that H⁺-ATP synthase-like molecules on the surface of HUVECs play an important role not only in extracellular ATP synthesis but also in the proliferation of HUVECs. The present results demonstrate that the use of small molecular H⁺-ATP synthase inhibitors targeting the F₁ catalytic domain may lead to significant advances in potential antiangiogenic cancer therapies.

This article has been cited by other articles:

				A. Lyly, S. K. Marjavaara, A. Kyttala, K. Uusi-Rauva, K. Luiro, O. Kopra, L. O. Martinez, K. Tanhuanpaa, N. Kalkkinen, A. Suomalainen, et al. Deficiency of the INCL protein Ppt1 results in changes in ectopic F1-ATP synthase and altered cholesterol metabolism Hum. Mol. Genet., May 15, 2008; 17(10): 1406 - 1417. [Abstract] [Full Text] [PDF]

				Y. Komi, O. Ohno, Y. Suzuki, M. Shimamura, K. Shimokado, K. Umezawa, and S. Kojima Inhibition of Tumor Angiogenesis by Targeting Endothelial Surface ATP Synthase with Sangivamycin Jpn. J. Clin. Oncol., November 1, 2007; 37(11): 867 - 873. [Abstract] [Full Text] [PDF]

				S. Erdmann, A. Ricken, C. Merkwitz, I. Struman, R. Castino, K. Hummitzsch, F. Gaunitz, C. Isidoro, J. Martial, and K. Spanel-Borowski The expression of prolactin and its cathepsin D-mediated cleavage in the bovine corpus luteum vary with the estrous cycle Am J Physiol Endocrinol Metab, November 1, 2007; 293(5): E1365 - E1377. [Abstract] [Full Text] [PDF]

				K. Yamamoto, N. Shimizu, S. Obi, S. Kumagaya, Y. Taketani, A. Kamiya, and J. Ando Involvement of cell surface ATP synthase in flow-induced ATP release by vascular endothelial cells Am J Physiol Heart Circ Physiol, September 1, 2007; 293(3): H1646 - H1653. [Abstract] [Full Text] [PDF]

				S. L. Chi, M. L. Wahl, Y. M. Mowery, S. Shan, S. Mukhopadhyay, S. C. Hilderbrand, D. J. Kenan, B. D. Lipes, C. E. Johnson, M. F. Marusich, et al. Angiostatin-Like Activity of a Monoclonal Antibody to the Catalytic Subunit of F1F0 ATP Synthase Cancer Res., May 15, 2007; 67(10): 4716 - 4724. [Abstract] [Full Text] [PDF]

				E. E. Quillen, G. C. Haslam, H. S. Samra, D. Amani-Taleshi, J. A. Knight, D. E. Wyatt, S. C. Bishop, K. K. Colvert, M. L. Richter, and P. A. Kitos Ectoadenylate Kinase and Plasma Membrane ATP Synthase Activities of Human Vascular Endothelial Cells J. Biol. Chem., July 28, 2006; 281(30): 20728 - 20737. [Abstract] [Full Text] [PDF]

				S. L. Chi and S. V. Pizzo Angiostatin Is Directly Cytotoxic to Tumor Cells at Low Extracellular pH: A Mechanism Dependent on Cell Surface-Associated ATP Synthase Cancer Res., January 15, 2006; 66(2): 875 - 882. [Abstract] [Full Text] [PDF]

				H. E. Burrell, B. Wlodarski, B. J. Foster, K. A. Buckley, G. R. Sharpe, J. M. Quayle, A. W. M. Simpson, and J. A. Gallagher Human Keratinocytes Release ATP and Utilize Three Mechanisms for Nucleotide Interconversion at the Cell Surface J. Biol. Chem., August 19, 2005; 280(33): 29667 - 29676. [Abstract] [Full Text] [PDF]

				J. Hu and M. M. Barr ATP-2 Interacts with the PLAT Domain of LOV-1 and Is Involved in Caenorhabditis elegans Polycystin Signaling Mol. Biol. Cell, February 1, 2005; 16(2): 458 - 469. [Abstract] [Full Text] [PDF]

				N. R. Burwick, M. L. Wahl, J. Fang, Z. Zhong, T. L. Moser, B. Li, R. A. Capaldi, D. J. Kenan, and S. V. Pizzo An Inhibitor of the F1 Subunit of ATP Synthase (IF1) Modulates the Activity of Angiostatin on the Endothelial Cell Surface J. Biol. Chem., January 21, 2005; 280(3): 1740 - 1745. [Abstract] [Full Text] [PDF]

				M. L. Wahl, T. L. Moser, and S. V. Pizzo Angiostatin and Anti-angiogenic Therapy in Human Disease Recent Prog. Horm. Res., January 1, 2004; 59(1): 73 - 104. [Abstract] [Full Text]