Nagata K, Sakagami H, Kondo H, Nozawa Y
Department of Molecular Pathobiochemistry, Gifu University School of Medicine, Japan.
Biochem Biophys Res Commun. 1995 Dec 26;217(3):1223-30. doi: 10.1006/bbrc.1995.2899.
The localization of gene expression of a low M(r) GTP-binding protein, ram p25, was examined throughout the entire brain of adult rat by in situ hybridization. ram-mRNA was expressed predominantly in anterior and intermediate lobes of pituitary gland (hypophysis), and weakly in hippocampal formation. In other parts of brain, no significant expression of ram-mRNA was detected, indicating that ram p25 may have an important role(s) in adenohypophysis. In 15-day-fetal rat, the mRNA was observed to be expressed strongly in the primitive anterior lobe of pituitary gland. The ram p25 was also detected in bovine pituitary gland by a specific antibody and was purified to near homogeneity by stepwise column chromatographies. Biochemical characterization of the purified ram p25 revealed that the profile of [35S]GTP gamma S-binding activity was almost the same as that of recombinant ram p25 expressed in E. coli.
通过原位杂交技术,在成年大鼠的整个大脑中检测了低分子量GTP结合蛋白ram p25的基因表达定位。ram - mRNA主要在垂体前叶和中叶表达,在海马结构中表达较弱。在大脑的其他部位,未检测到ram - mRNA的显著表达,这表明ram p25可能在腺垂体中发挥重要作用。在15日龄的胎鼠中,观察到该mRNA在垂体原始前叶中强烈表达。通过特异性抗体在牛垂体中也检测到了ram p25,并通过逐步柱色谱法将其纯化至接近均一性。对纯化的ram p25进行生化特性分析表明,[35S]GTPγS结合活性的图谱与在大肠杆菌中表达的重组ram p25几乎相同。