Purification of a noval protein phosphatase-1 binding protein, encoded by C9orf75.

• Main Authors: Yi-chun Tsai, 蔡宜君
• Other Authors: Hsien-bin Huang
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/46498257678322491038

碩士 === 國立中正大學 === 生命科學系暨分子生物研究所暨生物醫學研究 === 97 === Protein phosphatase-1 is one of the major serine/threonine protein phosphatases in eukaryotic cells. The catalytic subunit of protein phosphatase-1 (PP1) presents in cells as holoenzymes through association with a binding protein that targets the enzyme to specific subcellular compartments and regulates of the substrate functions, these substrate includ carbohydrate metabolism, protein synthesis, muscle contraction, transcription, cell cycle and neuronal signaling. Preview works in our lab have identified new PP1-binding as C9orf75. GST pull-down assay and co-immunoprecipitation have demonstrated that the protein product of C9orf75 is associated with PP1. The goal of my project is to understand the biochemical functions of C9orf75, prepared the recombinant thioredoxin-C9orf75 fusion protein (trx-C9orf75) from E.coli expression system by using Ni+2-Sepharose, Q-Sepharose and size-exclusion chromatographies. After Trx-tag of Trx-C9orf75 was cleaved by thrombin, the resulting C9orf75 was purified by gel-filtration.