Application). The biophysical parameters governing the interaction between the LDL-receptor related protein (LOP) and its ligands will be investigated. LRP is a scavenger receptor that is known to bind over twenty ligands, many of which have been found in proteinaceous plaques in the brains of persons suffering from Alzheimer's Disease (AD). In particular, this laboratory is interested in the LRP-mediated clearance of thrombin-PNI complexes, because the balance between thrombin and PN1 is upset in AD, and experiments have show l that thrombin acts to inhibit neurite outgrowth and promote synapse loss while PN1 reverses these effects. This laboratory has shown that clearance via LRP is the major route for clearance of thrombin-PN1 complexes by astrocyte cells, and it is suspected that disruption of LRP clearance may play a role in disruption of the balance between thrombin and PN1 in AD. LRP also clears ApoE, isoforms of which have been clearly linked genetically to AD, but the mechanism by which these isoforms either promote or suppress AD is not known. Because LRP has so many different ligands that could be causing many of the adverse effects seen in AD, these investigators propose to undertake careful biophysical experiments to understand ligand binding and ligand competition for LRP.
The aims of the proposed research are: (1) Express and characterize soluble fragments of the LRP extracellular subdomains. LRP has three repeated subdomains, each of which resembles the LDL receptor extracellular domain. These investigators will express each of these as well as smaller fragments of each in P. pastoris yeast. (2) Determine which LRP ligands compete for overlapping binding sites by ligand blotting. (3) Determine the smallest region of the LRP extracellular domain that bind to thrombin-PNlapoE, and PNII. (4) Characterize the kinetics of ligand binding using BIAcore. In particular, we will probe the role of hepara sulfate proteoglycans in the reaction. (5) Determine the structures of minimal fragments of LRP ligands bound to minimal fragments of the LRP extracellular domain.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Project (R01)
Project #
1R01AG017992-01
Application #
6092792
Study Section
Special Emphasis Panel (ZAG1-PKN-8 (J1))
Program Officer
Snyder, D Stephen
Project Start
2000-04-01
Project End
2004-03-31
Budget Start
2000-04-01
Budget End
2001-03-31
Support Year
1
Fiscal Year
2000
Total Cost
$244,568
Indirect Cost
Name
University of California San Diego
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
077758407
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Guttman, Miklos; Prieto, J Helena; Handel, Tracy M et al. (2010) Structure of the minimal interface between ApoE and LRP. J Mol Biol 398:306-19
Croy, Johnny E; Brandon, Theodore; Komives, Elizabeth A (2004) Two apolipoprotein E mimetic peptides, ApoE(130-149) and ApoE(141-155)2, bind to LRP1. Biochemistry 43:7328-35
Croy, Johnny E; Shin, William D; Knauer, Mary F et al. (2003) All three LDL receptor homology regions of the LDL receptor-related protein bind multiple ligands. Biochemistry 42:13049-57