Suzuki Y, Shimozawa N, Yajima S, Orii T, Yokota S, Tashiro Y, Osumi T, Hashimoto T
Department of Pediatrics, Gifu University School of Medicine, Japan.
Cell Struct Funct. 1992 Feb;17(1):1-8. doi: 10.1247/csf.17.1.
We investigated intracellular localization of peroxisomal proteins in fibroblasts from patients with Zellweger syndrome and neonatal adrenoleukodystrophy in whom peroxisomes were morphologically deficient or severely decreased. Indirect immunofluorescence staining revealed that catalase was mainly detected in the cytosol of fibroblasts from these patients, but a small amount of catalase was detected in granular pattern in a small percentage of cells. Double immunofluorescence staining revealed that catalase-containing particles in these patients also contained acyl-CoA oxidase and nonspecific lipid transfer protein. However, a 70 kD integral membrane protein and 3-ketoacyl-CoA thiolase were detected in all cells in granular pattern. Subcellular fractionation using digitonin after cell labeling revealed that a small amount of acyl-CoA oxidase and about half of thiolase in the precursor form were detected in the particulate fraction. These data suggest that the mechanisms of the transport and processing of catalase, acyl-CoA oxidase and nonspecific lipid transfer protein are different from those of the 70 kD integral membrane protein and 3-ketoacyl-CoA thiolase.
我们研究了 Zellweger 综合征和新生儿肾上腺脑白质营养不良患者成纤维细胞中过氧化物酶体蛋白的细胞内定位，这些患者的过氧化物酶体在形态上存在缺陷或数量严重减少。间接免疫荧光染色显示，过氧化氢酶主要在这些患者成纤维细胞的细胞质中检测到，但在一小部分细胞中以颗粒状模式检测到少量过氧化氢酶。双重免疫荧光染色显示，这些患者中含过氧化氢酶的颗粒也含有酰基辅酶 A 氧化酶和非特异性脂质转运蛋白。然而，在所有细胞中均以颗粒状模式检测到一种 70 kD 的整合膜蛋白和 3-酮酰基辅酶 A 硫解酶。细胞标记后使用洋地黄皂苷进行亚细胞分级分离显示，在颗粒部分检测到少量酰基辅酶 A 氧化酶和约一半前体形式的硫解酶。这些数据表明，过氧化氢酶、酰基辅酶 A 氧化酶和非特异性脂质转运蛋白的运输和加工机制与 70 kD 整合膜蛋白和 3-酮酰基辅酶 A 硫解酶的不同。
