Caveolin-1 is a negative regulator of endothelial nitric oxide synthase (eNOS) function. Indeed, caveolin-1 (-/-) mice exhibit enhanced endothelium-dependent relaxations in conduit vessels and exaggerated NO-dependent microvascular permeability in response to vascular endothelial growth factor (VEGF). We have shown that a cell-permeable, caveolin scaffolding domain (CSD) attenuates endothelium-dependent relaxations, and reduces vascular permeability in vivo. With this background in mind, we hypothesize that understanding the atomic interactions between eNOS and caveolin-1 will permit molecular dissection of the physiological and pathophysiological roles of caveolin-1 as a negative regulator of eNOS. To examine the regulation of this important interaction in more detail, we will:
Aim 1. Define the molecular interactions of eNOS with caveolin-1 at the atomic level. We will refine our structure of the CSD bound to eNOS and test the structural model using biophysical, biochemical and cell biological approaches. We will mutate key amino acid residues at the interface of eNOS and caveolin-1 and examine their effects on NOS activity and NO release;
Aim 2. Develop new cell-permeable peptides fused to the CSD based on structural information and test their effects in vivo. In preliminary results, we show a cell permeable version of CSD stereospecifically blocks endothelium-dependent relaxations, acute changes in vascular permeability and tumor permeability and progression in vivo. Moreover, we have isolated a small fragment of CSD that exerts biological activity in cells;
and Aim 3. Examine vascular function in transgenic mice that express mutant eNOS or caveolin-1. Based on structural information, we will define generate transgenic mice expressing wild-type or mutant CSD to directly test the molecular nature of this interaction. We will monitor systemic hemodynamics, vascular function, permeability, and tumor progression as physiological endpoints in these mice. ? ?

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL061371-07
Application #
6849274
Study Section
Pathology A Study Section (PTHA)
Program Officer
Goldman, Stephen
Project Start
1998-12-01
Project End
2009-01-31
Budget Start
2005-02-01
Budget End
2006-01-31
Support Year
7
Fiscal Year
2005
Total Cost
$408,750
Indirect Cost
Name
Yale University
Department
Pharmacology
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Luciano, Amelia K; Zhou, Wenping; Santana, Jeans M et al. (2018) CLOCK phosphorylation by AKT regulates its nuclear accumulation and circadian gene expression in peripheral tissues. J Biol Chem 293:9126-9136
Kuo, Andrew; Lee, Monica Y; Yang, Kui et al. (2018) Caveolin-1 regulates lipid droplet metabolism in endothelial cells via autocrine prostacyclin-stimulated, cAMP-mediated lipolysis. J Biol Chem 293:973-983
Luciano, Amelia K; Santana, Jeans M; Velazquez, Heino et al. (2017) Akt1 Controls the Timing and Amplitude of Vascular Circadian Gene Expression. J Biol Rhythms 32:212-221
Grabi?ska, Kariona A; Edani, Ban H; Park, Eon Joo et al. (2017) A conserved C-terminal RXG motif in the NgBR subunit of cis-prenyltransferase is critical for prenyltransferase activity. J Biol Chem 292:17351-17361
Hoffmann, Reuben; Grabi?ska, Kariona; Guan, Ziqiang et al. (2017) Long-Chain Polyprenols Promote Spore Wall Formation in Saccharomyces cerevisiae. Genetics 207:1371-1386
Kuo, Andrew; Lee, Monica Y; Sessa, William C (2017) Lipid Droplet Biogenesis and Function in the Endothelium. Circ Res 120:1289-1297
Kraehling, Jan R; Sessa, William C (2017) Contemporary Approaches to Modulating the Nitric Oxide-cGMP Pathway in Cardiovascular Disease. Circ Res 120:1174-1182
Grabi?ska, Kariona A; Park, Eon Joo; Sessa, William C (2016) cis-Prenyltransferase: New Insights into Protein Glycosylation, Rubber Synthesis, and Human Diseases. J Biol Chem 291:18582-90
Chamorro-Jorganes, Aránzazu; Lee, Monica Y; Araldi, Elisa et al. (2016) VEGF-Induced Expression of miR-17-92 Cluster in Endothelial Cells Is Mediated by ERK/ELK1 Activation and Regulates Angiogenesis. Circ Res 118:38-47
Park, Eon Joo; Grabi?ska, Kariona A; Guan, Ziqiang et al. (2016) NgBR is essential for endothelial cell glycosylation and vascular development. EMBO Rep 17:167-77

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