Diseases of the circulatory system constitute the leading cause of death in humans. Atherosclerosis involved in such diseases is initiated by attachment of monocytes to the arterial endothelial cells followed by migration through the arterial endothelial layer to the subendothelial space. this process is thought to involve monocyte chemotactic protein-1 (MCP-1) generated by endothelial and other cells in the vascular wall. Monocyte migration into injury sites is also involved in inflammation and wound healing. Understanding of the structure, function and regulation of production of MCP-1 could lead to new ways to deal with major diseases of man. To achieve such long-term goals, we propose the following specific objectives: 1. Determine structural features that mediate cell-type specificity of MCP-1 and the related neutrophil attractant protein (NAP-1/IL-8) using site-directed mutagenesis to produce structurally altered forms, bioassays to determine the activity of the mutants towards monocytes and neutrophils, and 2-D NMR to elucidate structural alterations in the mutants. 2. Clone a cDNA encoding the MCP-1 receptor, either using the likely similarity of this receptor to other chemotactic peptide receptors, or using labeled MCP-1 binding to COS cells expressing the cDNA clone, and attempts to engineer the receptor to elucidate the binding regions that confer specificity towards MCP-1 or IL-8. Knowledge of the structures of MCP-1 receptor, the regions of MCP that bind to the receptor, cloning of receptor and its structure and function will allow us to characterize the modulation of monocyte chemotaxis. This information would be important in designing treatments for atherosclerosis and other diseases that involve monocyte migration.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL048916-01A1
Application #
3368096
Study Section
Pathology A Study Section (PTHA)
Project Start
1993-04-01
Project End
1996-03-31
Budget Start
1993-04-01
Budget End
1994-03-31
Support Year
1
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Ohio State University
Department
Type
Schools of Engineering
DUNS #
098987217
City
Columbus
State
OH
Country
United States
Zip Code
43210
Randolph-Habecker, Julie-Randoph; Rahill, Brian; Torok-Storb, Beverly et al. (2002) The expression of the cytomegalovirus chemokine receptor homolog US28 sequesters biologically active CC chemokines and alters IL-8 production. Cytokine 19:37-46
Froberg, M K; Adams, A; Seacotte, N et al. (2001) Cytomegalovirus infection accelerates inflammation in vascular tissue overexpressing monocyte chemoattractant protein-1. Circ Res 89:1224-30
Moldovan, N I; Goldschmidt-Clermont, P J; Parker-Thornburg, J et al. (2000) Contribution of monocytes/macrophages to compensatory neovascularization: the drilling of metalloelastase-positive tunnels in ischemic myocardium. Circ Res 87:378-84
Crisman, J M; Elder, P J; Wilkie, N M et al. (1999) Identification of amino acids involved in the binding of hMIP-1 alpha to CC-CKR1, a MIP-1 alpha receptor found on neutrophils. Mol Cell Biochem 195:245-56
Beall, C J; Breckenridge, S M; Chakravarty, L et al. (1998) Expression of human monocyte chemoattractant protein-1 in the yeast Pichia pastoris. Protein Expr Purif 12:145-50
Chakravarty, L; Rogers, L; Quach, T et al. (1998) Lysine 58 and histidine 66 at the C-terminal alpha-helix of monocyte chemoattractant protein-1 are essential for glycosaminoglycan binding. J Biol Chem 273:29641-7
Kolattukudy, P E; Quach, T; Bergese, S et al. (1998) Myocarditis induced by targeted expression of the MCP-1 gene in murine cardiac muscle. Am J Pathol 152:101-11
Beall, C J; Mahajan, S; Kuhn, D E et al. (1996) Site-directed mutagenesis of monocyte chemoattractant protein-1 identifies two regions of the polypeptide essential for biological activity. Biochem J 313 ( Pt 2):633-40
Kuhn, D E; Beall, C J; Kolattukudy, P E (1995) The cytomegalovirus US28 protein binds multiple CC chemokines with high affinity. Biochem Biophys Res Commun 211:325-30
Li, Y S; Kolattukudy, P E (1994) Functional role of the cis-acting elements in human monocyte chemotactic protein-1 gene in the regulation of its expression by phorbol ester in human glioblastoma cells. Mol Cell Biochem 141:121-8