Chan Hsin-Tzu, Lee Tian-Ren, Huang Shun-Hong, Lee Hsiao-Yun, Sang Tzu-kang, Chan Hong-Lin, Lyu Ping-Chiang
Institute of Bioinformatics and Structural Biology & Department of Medical Sciences, National Tsing Hua University, Hsinchu, Taiwan.
Mol Biosyst. 2012 Jun;8(6):1730-41. doi: 10.1039/c2mb25037c. Epub 2012 Apr 5.
IBMPFD, Inclusion body myopathy associated with Paget's disease of bone and frontotemporal dementia, is a hereditary degenerative disorder due to single missense mutations in VCP (Valosin-Containing Protein). The mechanisms of how mutations of VCP lead to IBMPFD remain mysterious. Here we utilize two-dimensional difference gel electrophoresis (2D-DIGE) combined with mass spectrometry to study the IBMPFD disorder at the protein level. With this set-up, we are able to employ comparative proteomics to analyze IBMPFD disease using Drosophila melanogaster as our disease model organism. Head proteome of transgenic D. melanogaster expressing wild type VCP is compared, respectively, with the head proteome of transgenic mutant type VCPs that correspond to human IBMPFD disease alleles (TER94(A229E), TER94(R188Q), and TER94(R152H)). Of all the proteins identified, a significant fraction of proteins altered in TER94(A229E) and TER94(R188Q) mutants belong to the same functional categories, i.e. apoptosis and metabolism. Among these, Drosophila transferrin is observed to be significantly up-regulated in mutant flies expressing TER94(A229E). A knock-down experiment suggests that fly transferrin might be a potential modifier in IBMPFD disease. The molecular analysis of IBMPFD disease may benefit from the proteomics approach which combines the advantages of high throughput analysis and the focus on protein levels.
包涵体肌病伴骨Paget病和额颞叶痴呆（IBMPFD）是一种遗传性退行性疾病，由含缬酪肽蛋白（VCP）的单一位点错义突变引起。VCP突变导致IBMPFD的机制仍然不明。在此，我们利用二维差异凝胶电泳（2D-DIGE）结合质谱技术，在蛋白质水平研究IBMPFD疾病。通过这种方法，我们能够采用比较蛋白质组学，以黑腹果蝇作为疾病模式生物来分析IBMPFD疾病。将表达野生型VCP的转基因黑腹果蝇的头部蛋白质组，分别与对应人类IBMPFD疾病等位基因（TER94(A229E)、TER94(R188Q)和TER94(R152H)）的转基因突变型VCP的头部蛋白质组进行比较。在所有鉴定出的蛋白质中，TER94(A229E)和TER94(R188Q)突变体中发生改变的很大一部分蛋白质属于相同的功能类别，即细胞凋亡和代谢。其中，观察到在表达TER94(A229E)的突变果蝇中，果蝇转铁蛋白显著上调。一项敲低实验表明，果蝇转铁蛋白可能是IBMPFD疾病的一个潜在修饰因子。结合高通量分析优势和关注蛋白质水平的蛋白质组学方法，可能有助于对IBMPFD疾病进行分子分析。
