Study of cellular protein aggregates involved in the expression of harmful heavy metal toxicity.

2016 Fiscal Year Final Research Report

• Project/Area Number: 25340051
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Risk sciences of radiation and chemicals
• Research Institution: Jikei University School of Medicine
• Principal Investigator: TAKADA KOJI 東京慈恵会医科大学, 医学部, 教授 (30179452)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Keywords: 有害重金属 / カドミウム / メチル水銀 / タンパク凝集体 / ユビキチン / ポリユビキチン / ユビキチン化タンパク質

Outline of Final Research Achievements

To elucidate properties and significance of intracellular protein aggregates related to cytotoxicity of harmful heavy metals, in the analysis of aggregated components, we improved the purity of the aggregated protein fraction by reviewing the composition of cell extraction buffers. In addition, identification of aggregated components was carried out by a method based on shotgun analysis using high-resolution HPLC / Q-TOF. To study the formation mechanism of protein aggregation, we introduced the cell culture experiments using stable isotope-containing amino acids, and analyzed it in a time-dependent manner. Additionally, we improved the method of evaluation system of the protein aggregation using the hardly soluble polyubiquitinated proteins as an indication of cellular protein aggregation, and by this efficient method we verified and analyzed the effect of heavy metals on human epithelial cells.

Free Research Field

生化学・細胞生物学