ANALYSIS OF TEMPERATURE SENSITIVE IN PEROXISOME BIOGENESIS DISORDERS

(1)我们鉴定了31例日本过氧化酶体生物发生障碍(PBDS)患者，并在日本人的一条祖先染色体上发现了一种常见的突变。(2)我们鉴定了一个以PEX14为缺陷基因的新的PBD互补组。(3)为了阐明温度敏感现象的机制，我们利用纯化的重组蛋白，通过三维结构和远紫外圆二色谱分析了Pex13p-pex14p的相互作用，并利用患者的成纤维细胞进行了免疫荧光研究。(4)建立了超长链脂肪酸的气相色谱/质谱法分析过氧体疾病的筛查体系。(5)我们鉴定了一种过氧化酶体特异的Lon蛋白酶亚型，这是一种具有伴侣样活性的依赖于ATP的蛋白酶。(6)我们用过氧化体蛋白的双重免疫荧光染色证明了过氧化体在发育中的小鼠小脑浦肯野细胞和Bergmann神经胶质中的定位。

(1)We identified 31 Japanese patients with peroxisome biogenesis disorders (PBDs) and a common mutation arosed once on an ancestral chromosome in the Japanese population.(2)We identified a new complementation group of PBDs with PEX14 as the defective gene.(3)To clarify the mechanizm of temperature sensitive phenomenon, we analyze Pex13p-pex14p interaction by three dementional structure and far-UV circular dichroism spectra, using the purified recombinant proteins, and by the immunofluorescent studies, using the fibroblasts from the patients.(4)We established the screening system of peroxisomal disorders by gas chromatography/mass spectrometry anafysis of very long chain fatty acids, phytanic acid and plasmalogen.(5)We identified a peroxisome-specific isoform of Lon protease, an ATP-dependent protease with chaperone-like activities.(6)We demonstrated peroxisomal localization in Purkinje cells and Bergmann glia of the developing mouse cerebellum, using double immunofluorescent staining for peroxisomal proteins.