MEMBRANE TRAFFICKING IN MAMMALIAN CELLS

描述：在初步研究中，私家侦探提供了证据 高尔基体和内体膜的微管形成可能是由磷脂酶A2介导的。这是 基于一种利用牛重建高尔基体小管的体外实验 脑细胞液。一个~130kD的复合体因子，主要多肽为40 KD为分离株。多克隆抗血清对肾小管的免疫耗竭作用 来自BBC的活动，表明这个综合体确实在 体外培养。这一因素可能是PLA2是由PLA2的发现提供的 溴烯醇内酯等抑制剂抑制BFA诱导的肾小管上皮细胞高尔基体形成 体内和胞浆在体外诱导小管形成。一个类似的发现是 通过内吞体内获得的。此外，PLAP，一种激活PLA2的活性 刺激完整细胞的逆行运输，这种效应是 依赖于40kD管状因子的存在。最后， 含有磷脂酶活性的管状因子制剂 体外培养。P.I.提出了三个目标：(1)从分子上表征 40kD管状因子，并对其酶学、组织分布和组织分布进行了研究. 膜结合，(2)研究内吞体管形成及其影响因素， (3)膜内PLA2/膜管状结构的功能分析 贩卖人口。

DESCRIPTION: In preliminary studies, the P.I. has provided evidence that Golgi and endosome membrane tubulation may be mediated by PLA2. This is based on an in vitro assay that reconstitutes Golgi tubulation using bovine brain cytosol. A factor of ~130 kD complex with a major polypeptide of 40 kD was isolated. Polyclonal antiserum can immuno-deplete the tubulation activity from BBC, indicating that this complex is indeed functional in vitro. That this factor may be PLA2 is provided by the finding that PLA2 inhibitors such as bromoenol lactone inhibit BFA induced Golgi tubulation in vivo and cytosol induced tubulation in vitro. A similar finding was obtained with endosomes. Furthermore, PLAP, an activator PLA2 activity stimulates retrograde transport in intact cells, and this effect is dependent on the presence of the 40 kD tubulation factor. Finally, the tubulation factor preparation contained phospholipase activity as assayed in vitro. The P.I. proposed three aims: (1) to molecularly characterize the 40 kD tubulation factor and study its enzymology, tissue distribution and membrane association, (2) to study endosome tubulation and factors involved, (3) functional analysis of PLA2/ membrane tubulation in membrane trafficking.