Phosphoproteomic analysis of the human pituitary
- 257 Downloads
The pituitary is the central endocrine gland that regulates the functions of various target organs in the human body. Because of the pivotal regulatory role of the pituitary, it is essential to define on a global scale the components of the pituitary protein machinery, including a comprehensive characterization of the post-translational modifications of the pituitary proteins. Of particular interest is the examination of the phosphorylation status of the pituitary in health and disease. Towards the goal of global profiling of pituitary protein phosphorylation, we report here the application of the in-gel IEF-LC-MS/MS approach to the study of the pituitary phosphoproteome. The analytical strategy combined isoelectric focusing in immobilized pH gradient strips with immobilized metal ion affinity chromatography and mass spectrometry. With this method, a total of 50 phosphorylation sites were characterized in 26 proteins. Because the investigation involved primary tissue, the findings provide a direct glimpse into the phosphoprotein machinery operating within the human pituitary tissue microenvironment.
KeywordsHuman pituitary tissue Isoelectric focusing Mass spectrometry Phosphoproteome
Unable to display preview. Download preview PDF.
- 10.Gatlin CL, Kleemann GR, Hays LG, Link AJ, Yates JR III (1998) Protein identification at the low femtomole level from silver-stained gels using a new fritless electrospray interface for liquid chromatography-microspray and nanospray mass spectrometry. Anal Biochem 263:93–101PubMedCrossRefGoogle Scholar
- 22.Yoshizato H, Tanaka M, Nakai N, Nakao N, Nakashima K (2004) Growth hormone (GH)-stimulated insulin-like growth factor I gene expression is mediated by a tyrosine phosphorylation pathway depending on C-terminal region of human GH receptor in human GH receptor-expressing Ba/F3 cells. Endocrinology 145:214–220PubMedCrossRefGoogle Scholar