Protein expression and physiological function of Progesterone Receptor Membrane Component 1_Human

• Main Authors: Szu-Han Chou, 周思涵
• Other Authors: Kelvin H.-C. Chen
• Format: Others
• Language: zh-TW
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/23218108822510484825

碩士 === 國立屏東教育大學 === 化學生物系 === 99 === Progesterone Receptor Membrane Component 1 (PGRMC1) is a small membrane-associated protein, which widely found in eukaryotic cells. SDS-PAGE analysis show the protein band associated with the 28 kDa. Recently, many studies suggested that PGRMC1 and other membrane associated progesterone receptor proteins (MAPR) are directly bind to heme and cytochrome P450 superfamily members. The PGRMC1-CYP protein complex is playing an important role in cholesterol biosynthesis, steroid and drug metabolism. Furthermore, PGMRC1 associates with Epidermal Growth Factor Receptor (EGFP, a trans-membrane receptor tyrosine kinase to induce the cancer cell growth) and regulates Erlotinib (Target chemotherapy for cancer cell) sensitivity. In this study, we cloned the PGMRC1 gene by Saccharomyces cerevisiae, and employed 5-Aminolevulinic acid hydrochloride (ALA, the heme biosynthesis precursor) as the cell culture medium additive. The highly-heme loaded PGRMC1 has been harvested and purified by liquid chromatography successfully. In addition, the gas chromatography mass spectrometry (GC/MS) analysis indicated that the highly-heme loaded PGRMC1 is up-regulated the cytochrome P450 physiology activity. The influence of heme availability on PGRMC1 is evident. To further investigate the role of heme in cytochrome P450 regulation by PGRMC1, this study mutate several amino acid residues which decrease the heme binding capacity of PGRMC1 from previous reference, than observed the variety of ergosterol content.The result shows PGRMC1 and heme binding capacity will result in ergosterol content decreased, heme plays a key role in PGRMC1 regulates cytochrome P450 activity. Keywords: PGRMC1; cytochrome P450; sterol biosynthesis