Endothelial cell surface F1-F0 ATP synthase is active in ATP synthesis and is inhibited by angiostatin.
Angiostatin blocks tumor angiogenesis in vivo, almost certainly through its demonstrated ability to block endothelial cell migration and proliferation. Although the mechanism of angiostatin action remains unknown, identification of F(1)-F(O) ATP synthase as the major angiostatin-binding site on the endothelial cell surface suggests that ATP metabolism may play a role in the angiostatin response. Previous studies noting the presence of F(1) ATP synthase subunits on endothelial cells and certain cancer cells did not determine whether this enzyme was functional in ATP synthesis. We now demonstrate that all components of the F(1) ATP synthase catalytic core are present on the endothelial cell surface, where they colocalize into discrete punctate structures. The surface-associated enzyme is active in ATP synthesis as shown by dual-label TLC and bioluminescence assays. Both ATP synthase and ATPase activities of the enzyme are inhibited by angiostatin as well as by antibodies directed against the alpha- and beta-subunits of ATP synthase in cell-based and biochemical assays. Our data suggest that angiostatin inhibits vascularization by suppression of endothelial-surface ATP metabolism, which, in turn, may regulate vascular physiology by established mechanisms. We now have shown that antibodies directed against subunits of ATP synthase exhibit endothelial cell-inhibitory activities comparable to that of angiostatin, indicating that these antibodies function as angiostatin mimetics.
Moser
T L
TL
Department of Pathology and Duke University School of Nursing, Duke University Medical Center, Durham, NC 27710, USA.
Kenan
D J
DJ
Ashley
T A
TA
Roy
J A
JA
Goodman
M D
MD
Misra
U K
UK
Cheek
D J
DJ
Pizzo
S V
SV
D000255:Adenosine Triphosphate / Q000096:biosynthesis*; D043168:Angiostatins; D000818:Animals; D002384:Catalysis; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D004730:Endothelium, Vascular / Q000166:cytology / Q000201:enzymology*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D010446:Peptide Fragments / Q000494:pharmacology*; D010958:Plasminogen / Q000494:pharmacology*; D011487:Protein Conformation; D021122:Protein Subunits; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000502:physiology*
D004791:Enzyme Inhibitors; D010446:Peptide Fragments; D021122:Protein Subunits; D043168:Angiostatins; D000255:Adenosine Triphosphate; D010958:Plasminogen; D006180:Proton-Translocating ATPases