The goal of this renewal application (for years 18 to 22) is to define the function of the recently identified 60 kDa albumin-binding glycoprotein (gp60) on the endothelial cell membrane in mediating the transendothelial flux of albumin. We have purified gp60 from bovine pulmonary microvessel endothelial cell (BPMVEC) membranes and lungs. Cross-linking of gp60 using an anti-gp60 antibody (Ab) and a secondary Ab to activate gp60 increased the endocytosis and permeability of 125I-labeled albumin 2 to 3 fold in endothelial monolayers and pulmonary microvessels of rats. Gp60 activation also increased the uptake and transport of horseradish peroxidase (a fluid phase marker). Gp60 was shown to co-localize with caveolin-1, the principal structural protein of caveolae in endothelial cell membrane gp60 activates the tyrosine kinase signaling pathway that mediates the internalization and transcytosis of albumin. The proposed studies have the following aims: (1) to identify the albumin binding domains of gp60 and the mechanism of interaction of albumin with gp60 that activates endocytosis, (2) to determine in endothelial monolayers and pulmonary microvessels the function of gp60 activation in mediating transcytosis of albumin and fluid phase macromolecules, 93) to study interaction of gp60 with caveolin-1, and activation of tyrosine kinase signaling pathway in regulating albumin endocytosis and transcytosis in endothelial cells, and 94) to determine the function of gp60 in mediating albumin transcytosis by gp60 cloning and its expression. With the achievement of these aims, we will be able to provide new insights into this important mechanism of albumin permeability in pulmonary vascular endothelial cells.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL027016-19
Application #
2771237
Study Section
Lung Biology and Pathology Study Section (LBPA)
Project Start
1981-07-01
Project End
2002-08-31
Budget Start
1998-09-01
Budget End
1999-08-31
Support Year
19
Fiscal Year
1998
Total Cost
Indirect Cost
Name
University of Illinois at Chicago
Department
Pharmacology
Type
Schools of Medicine
DUNS #
121911077
City
Chicago
State
IL
Country
United States
Zip Code
60612
Minshall, Richard D; Vandenbroucke, Emily E; Holinstat, Michael et al. (2010) Role of protein kinase Czeta in thrombin-induced RhoA activation and inter-endothelial gap formation of human dermal microvessel endothelial cell monolayers. Microvasc Res 80:240-9
Fan, Jie; Frey, Randall S; Malik, Asrar B (2003) TLR4 signaling induces TLR2 expression in endothelial cells via neutrophil NADPH oxidase. J Clin Invest 112:1234-43
Fan, Jie; Malik, Asrar B (2003) Toll-like receptor-4 (TLR4) signaling augments chemokine-induced neutrophil migration by modulating cell surface expression of chemokine receptors. Nat Med 9:315-21
Ong, Evan S; Gao, Xiao-Pei; Xu, Ning et al. (2003) E. coli pneumonia induces CD18-independent airway neutrophil migration in the absence of increased lung vascular permeability. Am J Physiol Lung Cell Mol Physiol 285:L879-88
Mehta, Dolly; Tiruppathi, Chinnaswamy; Sandoval, Raudal et al. (2002) Modulatory role of focal adhesion kinase in regulating human pulmonary arterial endothelial barrier function. J Physiol 539:779-89
Frey, Randall S; Rahman, Arshad; Kefer, John C et al. (2002) PKCzeta regulates TNF-alpha-induced activation of NADPH oxidase in endothelial cells. Circ Res 90:1012-9
Fan, Jie; Frey, Randall S; Rahman, Arshad et al. (2002) Role of neutrophil NADPH oxidase in the mechanism of tumor necrosis factor-alpha -induced NF-kappa B activation and intercellular adhesion molecule-1 expression in endothelial cells. J Biol Chem 277:3404-11
Mehta, D; Rahman, A; Malik, A B (2001) Protein kinase C-alpha signals rho-guanine nucleotide dissociation inhibitor phosphorylation and rho activation and regulates the endothelial cell barrier function. J Biol Chem 276:22614-20
Kefer, J; Rahman, A; Anwar, K N et al. (2001) Decreased oxidant buffering impairs NF-kappaB activation and ICAM-1 transcription in endothelial cells. Shock 15:11-5
Rahman, A; Anwar, K N; Uddin, S et al. (2001) Protein kinase C-delta regulates thrombin-induced ICAM-1 gene expression in endothelial cells via activation of p38 mitogen-activated protein kinase. Mol Cell Biol 21:5554-65

Showing the most recent 10 out of 102 publications