PROTEIN PRODUCTION AND INTERACTION CORE (CORE D) The Protein Production and Interaction Core (Core D) will be led by W. Sean Davidson and will be located in his laboratory at the University of Cincinnati. His lab has extensive and documented experience in all proposed Core services. The Core is designed to provide a consistent and cost-effective set of high quality protein and lipoprotein reagents to PPG investigators in furtherance of the goals of each Project. Core D also provides evaluations of protein:protein interactions by developing and implementing state-of-the-art cross-linking approaches for application across the PPG projects as well as continued technology development with the aim of improving the resolution of structural determinations. The Core will be heavily used by each of the Projects. The goals of Core D are: 1. To express, purify and distribute recombinant apolipoproteins and their mutants produced in bacteria. 2. To express and distribute PON1, CETP, apoA-II and ABCA1 and their mutants produced in eukaryotic cells. 3. To isolate, characterize and distribute lipoproteins and apolipoproteins isolated from human plasma. 4. To generate, characterize and distribute reconstituted HDL lipoproteins. 5. To perform chemical cross-linking analyses on individual proteins, enzymes, transfer proteins and lipoprotein complexes in synergy with the HDL Quantitation Core (Core C). 6. To develop innovative new cross-linking approaches aimed at increasing protein structure resolution.

Public Health Relevance

CORE D The proposed research is relevant to public health because an increased understanding of the function of high density lipoproteins (HDL) will help guide the development of therapeutic strategies designed to raise plasma HDL for protection against cardiovascular disease, the # 1 killer in the U.S. Thus, the proposed research is relevant to the part of NIH's mission that pertains to developing fundamental knowledge for understanding the causes, prevention and eventually a cure for human diseases.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
1P01HL128203-01A1
Application #
9073919
Study Section
Special Emphasis Panel (HLBP (JH))
Project Start
2016-09-15
Project End
2021-06-30
Budget Start
2016-09-15
Budget End
2017-06-30
Support Year
1
Fiscal Year
2016
Total Cost
$119,137
Indirect Cost
$43,734
Name
Vanderbilt University Medical Center
Department
Type
DUNS #
079917897
City
Nashville
State
TN
Country
United States
Zip Code
37232
Lima, Diogo B; Melchior, John T; Morris, Jamie et al. (2018) Characterization of homodimer interfaces with cross-linking mass spectrometry and isotopically labeled proteins. Nat Protoc 13:431-458
Shao, Baohai; Heinecke, Jay W (2018) Quantifying HDL proteins by mass spectrometry: how many proteins are there and what are their functions? Expert Rev Proteomics 15:31-40
Cooke, Allison L; Morris, Jamie; Melchior, John T et al. (2018) A thumbwheel mechanism for APOA1 activation of LCAT activity in HDL. J Lipid Res 59:1244-1255
He, Yi; Kothari, Vishal; Bornfeldt, Karin E (2018) High-Density Lipoprotein Function in Cardiovascular Disease and Diabetes Mellitus. Arterioscler Thromb Vasc Biol 38:e10-e16
Pourmousa, Mohsen; Song, Hyun D; He, Yi et al. (2018) Tertiary structure of apolipoprotein A-I in nascent high-density lipoproteins. Proc Natl Acad Sci U S A 115:5163-5168
Melchior, John T; Walker, Ryan G; Cooke, Allison L et al. (2017) A consensus model of human apolipoprotein A-I in its monomeric and lipid-free state. Nat Struct Mol Biol 24:1093-1099
Melchior, John T; Street, Scott E; Andraski, Allison B et al. (2017) Apolipoprotein A-II alters the proteome of human lipoproteins and enhances cholesterol efflux from ABCA1. J Lipid Res 58:1374-1385