Overexpression, purification, and three dimensional stru

Investigators

Abstract

Autotaxin is a prometastatic enzyme central to the progression of various cancers, including ovarian carcinoma. Autotaxin is an extracellular protein which can either be attached to the cell surface or secreted into the extracellular milleu. The enzymatic domain is predicted to be an approximately 50 kD protein which becomes active upon homodimerization. This domain contains 6 cysteine residues, and is expressed as insoluble inclusion bodies in E.Coli expression systems. Our group proposes to refold and purify to homogeneity the ezymatic domain of autotaxin. Once autotaxin is produced in sufficient quantities, we propose to develop inhibitors to autotaxin by utilizing the NCI's resource in high throughput screening assays. In addition, we propose to solve the three dimensional structure of the protein using either x-ray crystallography or nuclear magnetic resonance to assist in structure related drug design. Overexpression, purification, and three dimensional structure determination of Autotaxin, a pro-metastatic stimulator of ovarian cancer.