Vitamin K is required for carboxylation of specific glutamic acid residues in proteins, converting them to gamma-carboxyglutamic acid (Gla). Gla residues in proteins exhibit specific calcium-binding and calcium-mediated phospholipid binding properties. Anticoagulants such as warfarin inhibit formation of Gla, thereby altering calcium-binding properties of Gla proteins. In addition to liver-derived blood coagulation factors and the well-characterized Gla protein of bone (osteocalcin), other normal tissues and tumor cell types produce Gla proteins of unknown function. In order to understand the mechanism by which warfarin anticoagulants inhibit tumor cell growth in vivo and in vitro, the basic biochemistry of tumor Gla proteins is under investigation. Model cell lines include mouse renal adenocarcinoma (RAG), human osteosarcoma (2T, MG63), and mouse melanoma (B16). The role of Gla proteins in proliferation, motility, and adhesion are under investigation. Warfarin inhibits growth in vitro and alters cellular thiol proteinase enzyme activity as well as generally depressing Gla-protein biosynthesis. Thus, the warfarin-sensitive target may be the cathepsin B-like thiol proteinase, which has been shown to correlate with malignancy in other studies. Recent studies have identified a precursor of this thiol proteinase that is secreted by tumor cells and whose activity is depressed 2- to 3-fold by warfarin. The molecular weight of the tumor cell procathepsin B is 44,000 and purification by affinity chromatography is in progress. (N)

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA037391-04
Application #
3175259
Study Section
Pathology B Study Section (PTHB)
Project Start
1983-01-01
Project End
1987-11-30
Budget Start
1985-12-01
Budget End
1986-11-30
Support Year
4
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Children's Hospital Boston
Department
Type
DUNS #
076593722
City
Boston
State
MA
Country
United States
Zip Code
02115
Hauschka, P V; Haroon, Y; Buchthal, S D et al. (1986) Vitamin K-dependent processes in tumor cells. Haemostasis 16:273-87
Hauschka, P V; Mavrakos, A E; Iafrati, M D et al. (1986) Growth factors in bone matrix. Isolation of multiple types by affinity chromatography on heparin-Sepharose. J Biol Chem 261:12665-74
Hauschka, P V (1986) Osteocalcin: the vitamin K-dependent Ca2+-binding protein of bone matrix. Haemostasis 16:258-72