Gel-Based Comparative Proteomics of Human Pituitary Adenoma Tissues

Gel-based comparative proteomics of human pituitary adenoma tissues 

Several critical features for the efficient extraction of proteins from a pituitary include (i) the pituitary control and adenoma tissues should be washed thoroughly with sodium chloride (0.9%) to remove any blood on the surface of the tissue (ii) due to the limited amount of the pituitary tissue (control and tumor ca. 0.5 g control ca. 15-80 mg adenoma), homogenization and lyophilization minimize any protein loss (iii) the pharmalyte in the protein solubilization buffer improved protein solubilization, stabilized the pi of proteins, and improved isoelectric focusing (lEF) with an IPG strip (iv) a combination of urea and thiourea improved protein solubilization and lEF [29] (v) repeated sonication improved protein solubilization  

The heterogeneity of human pituitary proteomes is a crucial factor that must be studied in order to accurately validate DEPs between pituitary adenomas and controls because the pituitary adenoma and control tissues are not from the same patient those different sources result in some uncontrolled experimental factors (gender, age, race) that occur within any comparative proteomics study of human pituitary adenomas. Therefore, the heterogeneity of a human pituitary proteome was studied as a function of three important factors—gender, age, and race [16]. 

A total of 30 high-resolution 2DGE gels from eight pituitary control tissues were used for a comparative analysis. A total of ca. 1000 protein spots were detected in each 2DGE map. Fifty-one differential spots (7 spots due to gender, 17 to age, 15 to race, and 12 to the co-effects of age and race) were found when pituitary 

Reference 2DGE map. One control pituitary and one adenoma tissue were analyzed with 2DGE and MS. For the control pituitary proteome [15], the 2D map contained 1094 protein spots a total of 62 spots, corresponding to 38 different proteins, were MS-characterized. For the pituitary adenoma tissue proteome [12], the 2D map contained ca. 1000 protein spots 135 protein spots that represented 111 proteins were MS-characterized. Those proteins correlated to different functional groups. The protein identification data were used to construct a Web-based reference database of the human pituitary (www.utmem.edu/proteomics). 

---