アクティブボード・２０１５年　４月
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研究発表を行った学会；
・第88回日本薬理学会年会
　２０１５年３月１８日〜２０日（名古屋）
タイトル；Characterization of the Intracellular Behavior of COL4A5 and Clarification of Molecular Mechanism of Alport Syndrome.
発表者；大町　紘平　氏
　　　（熊本大学　大学院薬学教育部　遺伝子機能応用学分野）
Abstract；
Alport syndrome (AS) is caused by mutation in COL4A3, COL4A4 or COL4A5, components of glomerular basement membrane (GBM). COL4A3/4/5 are secreted as heterotrimer. Restoring the normal COL4A3/4/5 network in the GBM is important for AS therapy; hence the need to investigate intracellular behavior of mutant COL4A5 such as degradation, heterotrimer formation and secretion. Here, we tried to establish AS cellular model and clarify the difference in degradation mechanism between wild type and mutant COL4A5 proteins.
We cloned and constructed human COL4A3/4/5 expression plasmids. These plasmids were transfected in HEK293 cells. We made mutant COL4A5 plasmids by site-directed mutagenesis. We are now investigating whether wild type and mutant COL4A5 are degraded by proteasome or lysosome by treating transfected cells with inhibitors. COL4A5 protein stability is also checked by chase experiments.
We made a novel AS cellular model, which expresses human COL4A3/4/5 proteins. This model is an important tool to investigate AS phenotype in molecular level such as protein stability, degradation mechanism, and heterotrimer formation of wild type or mutant COL4A5.