Heikoop J C, Van den Berg M, Strijland A, Weijers P J, Schutgens R B, Just W W, Wanders R J, Tager J M
E.C. Slater Institute for Biochemical Research, University of Amsterdam, Academic Medical Center, The Netherlands.
Biochim Biophys Acta. 1991 Jul 26;1097(1):62-70. doi: 10.1016/0925-4439(91)90025-5.
Rhizomelic Chondrodysplasia Punctata (RCDP) is an autosomal recessive disorder in which plasmalogen biosynthesis and phytanate catabolism are impaired. Peroxisomal structure and the intracellular localization of catalase, the 69 kDa peroxisomal integral membrane protein (PMP), and 3-oxoacyl-CoA thiolase were studied in cultured skin fibroblasts from control subjects and patients with RCDP. A punctate fluorescence pattern characteristic for peroxisomes was seen in control cells incubated with either anti-(catalase), anti-(69 kDa PMP) or anti-(3-oxoacyl-CoA thiolase). Incubation of mutant cells with anti-(catalase) or anti-(69 kDa PMP) resulted in the same pattern. However, when RCDP fibroblasts were incubated with a monoclonal anti-(3-oxoacyl-CoA thiolase) antibody no punctate fluorescence could be observed. Cryosections from control and RCDP cells were examined by electron microscopy using double immunogold labelling. RCDP fibroblasts contained structures indistinguishable from control peroxisomes, the membranes reacting with anti-(69 kDa PMP) and the matrix with anti-(catalase). However, the matrix of RCDP peroxisomes, unlike control peroxisomes, did not react with anti-(3-oxoacyl-CoA thiolase). We conclude that RCDP fibroblasts contain regularly shaped peroxisomes, comparable to control peroxisomes in number as well as in content of catalase and 69 kDa PMP. However, in RCDP peroxisomes the amount of 3-oxoacyl-CoA thiolase protein proved to be below the limit of detection.
肢根型点状软骨发育不良（RCDP）是一种常染色体隐性疾病，其中缩醛磷脂生物合成和植烷酸分解代谢受损。研究了来自对照受试者和RCDP患者的培养皮肤成纤维细胞中的过氧化物酶体结构以及过氧化氢酶、69 kDa过氧化物酶体整合膜蛋白（PMP）和3-氧代酰基辅酶A硫解酶的细胞内定位。在用抗（过氧化氢酶）、抗（69 kDa PMP）或抗（3-氧代酰基辅酶A硫解酶）孵育的对照细胞中，观察到过氧化物酶体特有的点状荧光模式。用抗（过氧化氢酶）或抗（69 kDa PMP）孵育突变细胞产生相同的模式。然而，当RCDP成纤维细胞与单克隆抗（3-氧代酰基辅酶A硫解酶）抗体孵育时，则观察不到点状荧光。使用双重免疫金标记通过电子显微镜检查对照细胞和RCDP细胞的冷冻切片。RCDP成纤维细胞含有与对照过氧化物酶体无法区分的结构，膜与抗（69 kDa PMP）反应，基质与抗（过氧化氢酶）反应。然而，与对照过氧化物酶体不同，RCDP过氧化物酶体基质不与抗（3-氧代酰基辅酶A硫解酶）反应。我们得出结论，RCDP成纤维细胞含有形状规则的过氧化物酶体，其数量以及过氧化氢酶和69 kDa PMP含量与对照过氧化物酶体相当。然而，在RCDP过氧化物酶体中，3-氧代酰基辅酶A硫解酶蛋白量被证明低于检测限。
