Presence of peroxisomal membrane proteins in liver and fibroblasts from patients with the Zellweger syndrome and related disorders: evidence for the existence of peroxisomal ghosts.
Presence of peroxisomal membrane proteins in liver and fibroblasts from patients with the Zellweger syndrome and related disorders: evidence for the existence of peroxisomal ghosts.
复制标题
齐薇格综合征及相关疾病患者的肝脏和成纤维细胞中存在过氧化物酶体膜蛋白：过氧化物酶体鬼影存在的证据。
DOI：
--
复制
发表时间：
1989
影响因子：
6.6
通讯作者：
A. Schram
中科院分区：
文献类型：
--
作者：
E. Wiemer;S. Brul;Just Ww;R. Vandriel;E. Brouwer;M. Vandenberg;Weijers Pj;R. Schutgens;H. Vandenbosch;A. Schram
过氧化物酶体的存在和细胞内定位的膜蛋白（PMP）进行了研究，在肝脏和培养的皮肤成纤维细胞从对照组和患者的Zellweger综合征和相关疾病，其中过氧化物酶体几乎是不存在的。免疫印迹实验表明，22，36和69 kDa的PMPs的存在，并局限于在对照组的肝脏和肝脏中的Zellweger患者的膜部分。22和36 kDa PMPs在患者肝脏中的含量显著低于对照肝脏。在来自一个Zellweger的肝脏中发现了减少量的69 kDa PMP，但在来自另一个Zellweger的肝脏中未发现。用间接免疫荧光法研究了过氧化氢酶和69 kDa PMP在成纤维细胞中的亚细胞定位。在与抗-（过氧化氢酶）或抗-（69 kDa PMP）孵育的对照细胞中观察到特征性点状荧光。孵育的突变体细胞与抗（过氧化氢酶）导致在一个弥漫的荧光，而与抗（69 kDa PMP）荧光颗粒可视化，在某些细胞系中，是更大的和更少的数量比对照细胞。使用免疫金标记通过电子显微镜检查对照和突变细胞的冷冻切片。对照细胞含有小的结构组成的一个单一的膜封闭一个均匀的矩阵;膜与抗（69 kDa PMP）和矩阵与抗（过氧化氢酶）。突变体细胞系含有球形或椭球形结构，其膜与抗（69 kDa PMP）反应;未观察到标记与抗（过氧化氢酶）。我们得出结论，过氧化物酶体鬼，其膜含有69 kDa的PMP，目前在过氧化物酶体缺陷的细胞系从所有的互补组研究。
The presence and intracellular localization of peroxisomal integral membrane proteins (PMP) were investigated in liver and cultured skin fibroblasts from control subjects and patients with the Zellweger syndrome and related disorders in which peroxisomes are virtually absent. Immunoblotting experiments showed that 22, 36 and 69 kDa PMPs were present and were confined to the membranous fraction both in the control liver and in the livers from the Zellweger patients. The 22 and 36 kDa PMPs were present in significantly lower amounts in the patients' livers than in the control liver. A reduced amount of the 69 kDa PMP was found in liver from one Zellweger but not in liver from another. The subcellular localization in fibroblasts of catalase and the 69 kDa PMP was studied by indirect immunofluorescence. A characteristic punctate fluorescence was seen in control cells incubated with either anti-(catalase) or with anti-(69 kDa PMP). Incubation of mutant cells with anti-(catalase) resulted in a diffuse fluorescence, whereas with anti-(69 kDa PMP) fluorescent particles were visualized which, in some cell lines, were larger and fewer in number than in control cells. Cryosections of control and mutant cells were examined by electron microscopy using immunogold labeling. Control cells contained small structures consisting of a single membrane enclosing a homogeneous matrix; the membranes reacted with anti-(69 kDa PMP) and the matrix with anti-(catalase). The mutant cell lines contained spherical or ellipsoidal structures whose membranes reacted with anti-(69 kDa PMP); no labeling was observed with anti-(catalase). We conclude that peroxisomal ghosts, the membranes of which contain the 69 kDa PMP, are present in peroxisome-deficient cell lines from all complementation groups studied so far.