title: Orai3 Surface Accumulation and Calcium Entry Evoked by Vascular Endothelial Growth Factor.
abstract: OBJECTIVE
  Vascular endothelial growth factor (VEGF) acts, in part, by triggering calcium ion (Ca(2+)) entry. Here, we sought understanding of a Synta66-resistant Ca(2+) entry pathway activated by VEGF.
  
  
  RESULTS
  Measurement of intracellular Ca(2+) in human umbilical vein endothelial cells detected a Synta66-resistant component of VEGF-activated Ca(2+) entry that occurred within 2 minutes after VEGF exposure. Knockdown of the channel-forming protein Orai3 suppressed this Ca(2+) entry. Similar effects occurred in 3 further types of human endothelial cell. Orai3 knockdown was inhibitory for VEGF-dependent endothelial tube formation in Matrigel in vitro and in vivo in the mouse. Unexpectedly, immunofluorescence and biotinylation experiments showed that Orai3 was not at the surface membrane unless VEGF was applied, after which it accumulated in the membrane within 2 minutes. The signaling pathway coupling VEGF to the effect on Orai3 involved activation of phospholipase Cγ1, Ca(2+) release, cytosolic group IV phospholipase A2α, arachidonic acid production, and, in part, microsomal glutathione S-transferase 2, an enzyme which catalyses the formation of leukotriene C4 from arachidonic acid. Shear stress reduced microsomal glutathione S-transferase 2 expression while inducing expression of leukotriene C4 synthase, suggesting reciprocal regulation of leukotriene C4-synthesizing enzymes and greater role of microsomal glutathione S-transferase 2 in low shear stress.
  
  
  CONCLUSIONS
  VEGF signaling via arachidonic acid and arachidonic acid metabolism causes Orai3 to accumulate at the cell surface to mediate Ca(2+) entry and downstream endothelial cell remodeling.
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Alexander-Francisco
authors.initials: AF
authors.lastname: Bruns
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Bing
authors.initials: B
authors.lastname: Hou
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Baptiste
authors.initials: B
authors.lastname: Rode
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Webster
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Marc A
authors.initials: MA
authors.lastname: Bailey
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Hollie L
authors.initials: HL
authors.lastname: Appleby
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Nicholas K
authors.initials: NK
authors.lastname: Moss
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Judith E
authors.initials: JE
authors.lastname: Ritchie
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Nadira Y
authors.initials: NY
authors.lastname: Yuldasheva
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Sarka
authors.initials: S
authors.lastname: Tumova
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Matthew
authors.initials: M
authors.lastname: Quinney
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Lynn
authors.initials: L
authors.lastname: McKeown
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Hilary
authors.initials: H
authors.lastname: Taylor
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: K Raj
authors.initials: KR
authors.lastname: Prasad
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Dermot
authors.initials: D
authors.lastname: Burke
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: David
authors.initials: D
authors.lastname: O'Regan
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Karen E
authors.initials: KE
authors.lastname: Porter
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Richard
authors.initials: R
authors.lastname: Foster
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.).
authors.forename: Mark T
authors.initials: MT
authors.lastname: Kearney
authors.affiliation: From the Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine (J.L., A.-F.B., B.H., B.R., P.J.W., M.A.B., H.L.A., N.K.M., J.E.R., N.Y.Y., S.T., M.Q., L.M., H.T., K.E.P., D.J.B.) and School of Chemistry (R.F.), University of Leeds, Leeds, United Kingdom; Departments of Hepatobiliary and Transplant Surgery (K.R.P.) and Colorectal Surgery (D.B.), St. James's University Hospital, Leeds, United Kingdom; and Yorkshire Heart Centre, Leeds General Infirmary, Leeds, United Kingdom (D.O.R.). d.j.beech@leeds.ac.uk.
authors.forename: David J
authors.initials: DJ
authors.lastname: Beech
mesh_terms: D000818:Animals; D050197:Atherosclerosis / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000096:biosynthesis / Q000235:genetics*; D002465:Cell Movement; D002478:Cells, Cultured; D004195:Disease Models, Animal; D005786:Gene Expression Regulation; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008819:Mice, Nude; D012313:RNA / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism; D066253:Vascular Remodeling / Q000235:genetics*
chemical_list: D015220:Calcium Channels; C520432:Orai3 protein, mouse; D042461:Vascular Endothelial Growth Factor A; D012313:RNA; D002118:Calcium
keywords: Orai3 protein; calcium; cytosol; endothelial cells; vascular endothelial growth factor A