Alexson S E, Fujiki Y, Shio H, Lazarow P B
J Cell Biol. 1985 Jul;101(1):294-304. doi: 10.1083/jcb.101.1.294.
Rat liver peroxisomes were subjected to a variety of procedures intended to partially disassemble or damage them; the effects were analyzed by recentrifugation into sucrose gradients, enzyme analyses, electron microscopy, and SDS PAGE. Freezing and thawing or mild sonication released some matrix proteins and produced apparently intact peroxisomal "ghosts" with crystalloid cores and some fuzzy fibrillar content. Vigorous sonication broke open the peroxisomes but the membranes remained associated with cores and fibrillar and amorphous matrix material. The density of both ghosts and more severely damaged peroxisomes was approximately 1.23. Pyrophosphate (pH 9) treatment solubilized the fibrillar content, yielding ghosts that were empty except for cores. Some matrix proteins such as catalase and thiolase readily leak from peroxisomes. Other proteins were identified that remain in mechanically damaged peroxisomes but are neither core nor membrane proteins because they can be released by pyrophosphate treatment. These constitute a class of poorly soluble matrix proteins that appear to correspond to the fibrillar material observed morphologically. All of the peroxisomal beta-oxidation enzymes are located in the matrix, but they vary greatly in how easily they leak out. Palmitoyl coenzyme A synthetase is in the membrane, based on its co-distribution with the 22-kilodalton integral membrane polypeptide.
对大鼠肝脏过氧化物酶体进行了各种旨在使其部分解体或受损的操作；通过再次离心至蔗糖梯度、酶分析、电子显微镜和SDS-PAGE对其效果进行了分析。冷冻和解冻或温和超声处理释放了一些基质蛋白，并产生了明显完整的过氧化物酶体“空壳”，其具有晶体核心和一些模糊的纤维状内含物。剧烈超声处理使过氧化物酶体破裂，但膜仍与核心以及纤维状和无定形基质物质相关联。空壳和受损更严重的过氧化物酶体的密度约为1.23。焦磷酸（pH 9）处理使纤维状内含物溶解，产生除核心外为空的空壳。一些基质蛋白，如过氧化氢酶和硫解酶，很容易从过氧化物酶体中泄漏出来。还鉴定出了其他一些蛋白，它们保留在机械损伤的过氧化物酶体中，但既不是核心蛋白也不是膜蛋白，因为它们可通过焦磷酸处理释放出来。这些构成了一类难溶性基质蛋白，似乎与形态学上观察到的纤维状物质相对应。所有过氧化物酶体β氧化酶都位于基质中，但它们泄漏的难易程度差异很大。基于棕榈酰辅酶A合成酶与22千道尔顿整合膜多肽的共分布，其位于膜中。
