Vitamin K-dependent proteins have been shown to play a major role in vertebrates in blood coagulation and its regulation, as well as in bone resorption and tissue mineralization. The vitamin K-dependent proteins contain gamma carboxyglutamic acid residues that are generated in a post-translation event within the lumen of the endoplasmic reticulum. This reaction is catalyzed by a gamma glutamyl carboxylase that is widely distributed in numerous vertebrate and non-vertebrate tissues. This investigation will focus on gaining insight into the biological role of four novel vitamin K-dependent proteins that are single transmembrane proteins. These proteins range in size from 17 kDa to 25 kDa and contain from nine to thirteen gamma- carboxyglutamic acid residues. They have been called PRGP1, PRGP2, TMG3, and TMG4. Genes for two of these transmembrane proteins, PRGP1 and TMG3 are located on the X chromosome, while genes for PRGP2 and TMG4 are found on chromosome 19 and 11, respectively. Studies with transfected mammalian cells have now shown that they require vitamin K for their post-translational modification in a reaction that is inhibited by warfarin. The carboxyl region of the four transmembrane proteins bind to Nedd4-1ike proteins, suggesting that they may play a role in the ubiquitin cycle. Nedd4-1ike proteins have a C2 domain, two to four WW domains, and a HECT domain that functions as an E3 ubiquitin ligase in the ubiquitin cycle. The mechanism and specificity of these reactions will be examined in detail in these investigations.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL071599-05
Application #
7163000
Study Section
Nutrition Study Section (NTN)
Program Officer
Sarkar, Rita
Project Start
2002-12-15
Project End
2008-05-30
Budget Start
2006-12-01
Budget End
2008-05-30
Support Year
5
Fiscal Year
2007
Total Cost
$287,488
Indirect Cost
Name
University of Washington
Department
Biochemistry
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Bridenbaugh, Eric A (2012) Isolation and preparation of RNA from rat blood and lymphatic microvessels for use in microarray analysis. Methods Mol Biol 843:265-89
Mize, Gregory J; Harris, Jeff E; Takayama, Thomas K et al. (2008) Regulated expression of active biotinylated G-protein coupled receptors in mammalian cells. Protein Expr Purif 57:280-9
Kulman, John D; Satake, Masanobu; Harris, Jeff E (2007) A versatile system for site-specific enzymatic biotinylation and regulated expression of proteins in cultured mammalian cells. Protein Expr Purif 52:320-8
Kulman, John D; Harris, Jeff E; Xie, Ling et al. (2007) Proline-rich Gla protein 2 is a cell-surface vitamin K-dependent protein that binds to the transcriptional coactivator Yes-associated protein. Proc Natl Acad Sci U S A 104:8767-72
Davie, Earl W; Kulman, John D (2006) An overview of the structure and function of thrombin. Semin Thromb Hemost 32 Suppl 1:3-15
Kulman, John D; Harris, Jeff E; Nakazawa, Noriko et al. (2006) Vitamin K-dependent proteins in Ciona intestinalis, a basal chordate lacking a blood coagulation cascade. Proc Natl Acad Sci U S A 103:15794-9