G protein-coupled receptors mediate coronary flow- and agonist-induced responses via lectin-oligosaccharide interactions
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Blood flow acts parallel to the coronary luminal endothelial surface layer (LESL) and modulates multiple parenchymal functions via the release of paracrine agents. Evidence indicates that the LESL may be a flow-sensing organelle and that perhaps through flow-induced lectin (L)·oligosaccharide (O) complex formation (L·O) participates in this process. LESL integrins and selectins are both lectinic and flow sensitive, but the L properties of flow-sensitive G protein-coupled receptors (GPCRs) are unknown. Therefore, we investigated the presence of L in the LESL and hypothesized that if flow-sensitive GPCRs are L, flow and O will determine their response to receptor activation. The LESL protein fraction isolated from guinea pig hearts was passed through an affinity chromatography column made of three sugars, mannose, galactose, and N-acetylglucosamine, and the lectinic fraction was eluted. Immune dot blot was used to identify L proteins in the LESL fraction. Our results indicate the following. 1) Two-dimensional SDS-PAGE (2D-SDS-PAGE) of the LESL lectinic fraction revealed at least 167 Ls. 2) Among these Ls, we identified three selectins and the GPCRs: angiotensin II, bradykinin (B2-R), adenosine A1 and A2, prolactin, endothelin, α1-adrenergic (α1A-R), thromboxane A2, β1-adrenergic, β3-adrenergic, and insulin receptors; the first six GPCRs are known to be flow sensitive. 3) The amplitude of receptor-induced vascular responses by α1A-R and B2-R activation (phenylephrine or bradykinin, respectively) was a function of flow and O (hyaluronidate). Our results support a novel mechanism of GPCR-mediated responses to flow via L·O interaction. © 2014 the American Physiological Society.
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Flow sensitivity requirements; G protein-coupled receptors; Lectins and sugars of glycocalyx; Mechanosensors; Regulatory extracellular matrix adenosine A1 receptor; adenosine A2 receptor; alpha 1 adrenergic receptor; angiotensin II; beta 1 adrenergic receptor; beta 3 adrenergic receptor; bradykinin receptor; carbohydrate; CD31 antigen; dihydrolipoamide dehydrogenase; endothelin; G protein coupled receptor; galactose; insulin receptor; integrin; intercellular adhesion molecule 1; lectin; mannose; n acetylglucosamine; oligosaccharide; prolactin; recombinant hyaluronidase; selectin; thromboxane A2 receptor; vascular cell adhesion molecule 1; cardiovascular agent; G protein coupled receptor; galectin; lectin; ligand; mannose binding lectin; n acetylglucosamine receptor; oligosaccharide; affinity chromatography; animal experiment; animal tissue; article; binding affinity; blood vessel reactivity; cell organelle; complex formation; concentration response; coronary artery blood flow; coronary luminal endothelial surface layer; endothelium cell; enzyme activation; glycocalyx; guinea pig; nonhuman; polyacrylamide gel electrophoresis; priority journal; protein binding; protein interaction; Western blotting; animal; biological model; blood flow; coronary blood vessel; dose response; drug effect; drug potentiation; flow sensitivity requirements; G protein-coupled receptors; lectins and sugars of glycocalyx; mechanosensors; mechanotransduction; metabolism; methodology; proteomics; regulatory extracellular matrix; time; vascular endothelium; flow sensitivity requirements; G protein-coupled receptors; lectins and sugars of glycocalyx; mechanosensors; regulatory extracellular matrix; Acetylglucosamine; Animals; Cardiovascular Agents; Chromatography, Affinity; Coronary Circulation; Coronary Vessels; Dose-Response Relationship, Drug; Electrophoresis, Polyacrylamide Gel; Endothelium, Vascular; Galactose; Galectins; Glycocalyx; Guinea Pigs; Lectins; Ligands; Mannose; Mannose-Binding Lectin; Mechanotransduction, Cellular; Models, Cardiovascular; Oligosaccharides; Proteomics; Receptors, G-Protein-Coupled; Receptors, N-Acetylglucosamine; Regional Blood Flow; Time Factors
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