Characterization of a basement membrane protein (BM180) using capillary electrophoresis

Coeliac disease (gluten intolerance) is a genetically based autoimmune disease that becomes evident following ingestion of cereal prolamins such as the wheat gliadins. The process of this disease is not yet thoroughly understood. Purification of basement membrane protein-180 (BM180) (a basement membrane protein with a potential autoantigen role) and multiple analysis of the purified protein can lead to a better understanding of this disorder, which affects millions of people worldwide. Our preliminary work on the purification and characterization of this protein is presented in this paper. BM180 proteins were expressed in mouse EHS (Engelberth-Holm-Swarm) tumor cells. A crude purification process (see "Methods") was performed and the purified fractions were analyzed by capillary gel electrophoresis (CGE) for determination of the apparent molecular weight of the protein components in each fraction. The fractions, which contained compounds of the expected molecular weight, were further analyzed using a capillary zone electrophoresis (CZE) method developed for the routine analysis of wheat gliadins. Using the two CE methods, we were able to compare BM180 with certain gliadin fractions. Additional information on the protein stability was also obtained.