Proteome Analysis Of Mammal Plasma Membrane And Breast Carcinomatous Proteome

Plasma membrane (PM) constitutes the interface between cells and their external environment. The proteins embedded in this membrane work as the "doorbells" and "doorways" of the cells, and are the targets of about 70% of all known drugs. For these reasons, proteomic analysis of the plasma membrane is very important for theoretical and applied cytology and pharmacology. However, due to the low abundance, high hydrophobicity and the evidently complexity of PM proteins, it is very challenging to study.First, we used mouse/rat liver as material, and looked for the methods for PM proteome research. The liver is one of the most important organs in the body, probably second only to the brain in organ complexity. Therefore, proteomic analysis of liver PM will be helpful to understand these physiology activities and treat liver disease. We isolated PM from mouse liver by sucrose density gradient centrifugation. The optimized lysis buffer containing 4% CHAPS and 1% NP-40 was obtained. The methods for integral membrane protein enrichmentwere analyzed and better methods----chloroform/methanolextraction and sodium carbonate treatment were found. The extracted PM proteins were separated by 2DE and SDS-PAGE, and submitted to MS identification. In all, 175 non-redundant proteins were identified, of which 9% proteins with unknown function. Through prediction, four unknown proteins were found to be related with limb-girdle muscular dystrophy 2B, and so on. Second, in order to characterize low copy integral membrane proteins. Integral plasma membrane proteins were enriched through the treatment with 0. 1M Na₂CO₃, chloroform/methanol...