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首页> 外文期刊>Biochimica et biophysica acta. Biomembranes >Characterization of the first planctomycetal outer membrane protein identifies a channel in the outer membrane of the anammox bacterium Kuenenia stuttgartiensis
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Characterization of the first planctomycetal outer membrane protein identifies a channel in the outer membrane of the anammox bacterium Kuenenia stuttgartiensis

机译：第一个平分缩醛外膜蛋白的表征鉴定了厌氧细菌KueneniaStuttgartiensis的外膜中的通道

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摘要

Planctomycetes are a bacterial phylum known for their complex intracellular compartmentalization. While most Planctomycetes have two compartments, the anaerobic ammonium oxidizing (anammox) bacteria contain three membrane-enclosed compartments. In contrast to a long-standing consensus, recent insights suggested the outermost Planctomycete membrane to be similar to a Gram-negative outer membrane (OM). One characteristic component that differentiates OMs from cytoplasmic membranes (CMs) is the presence of outer membrane proteins (OMPs) featuring a beta-barrel structure that facilitates passage of molecules through the OM. Although proteomic and genomic evidence suggested the presence of OMPs in several Planctomycetes, no experimental verification existed of the pore-forming function and localization of these proteins in the outermost membrane of these exceptional microorganisms. Here, we show via lipid bilayer assays that at least two typical OMP-like channel-forming proteins are present in membrane preparations of the anammox bacterium Kuenenia stuttgartiensis. One of these channel-forming proteins, the highly abundant putative OMP Kustd1878, was purified to homogeneity. Analysis of the channel characteristics via lipid bilayer assays showed that Kustd1878 forms a moderately cation-selective channel with a high current noise and an average single-channel conductance of about 170-190 pS in 1 M KC1. Antibodies were raised against the purified protein and immunogold localization indicated Kustd1878 to be present in the outermost membrane. Therefore, this work clearly demonstrates the presence of OMPs in anammox Planctomycetes and thus firmly adds to the emerging view that Planctomycetes have a Gram-negative cell envelope.
机译：Planctomycetes是一种已知其复杂的细胞内隔室化的细菌字节。虽然大多数pancteromycetes有两个隔室，但厌氧铵氧化（厌氧毒素）细菌含有三个膜封闭的隔室。与长期共识相比，最近的见解表明最外面的平分蜱膜与革兰氏阴性外膜（OM）相似。将OMS与细胞质膜（CMS）区分开的一个特征组分是存在外膜蛋白（OMP）的存在，其具有β-镜筒结构，其有助于分子通过OM通过。尽管蛋白质组学和基因组证据表明，在几种planctomycetes中存在OMP的存在，但在这些特殊微生物的最外膜中没有存在孔形成功能和这些蛋白质的定位的实验验证。在这里，我们通过脂质双层测定显示，至少两个典型的OMP样沟道形成蛋白质存在于厌氧细菌Kuenenia Stuttgartiensis的膜制剂中。将这些沟道形成蛋白质中的一种，高度充裕的投资KustD1878纯化为均匀性。通过脂质双层测定分析通道特性，显示Kustd1878在1M KC1中形成具有高电流噪声的中等阳离子选择性通道和约170-190ps的平均单通道电导。针对纯化的蛋白质升高抗体，并在最外膜中存在显着的kustd1878。因此，这项工作清楚地证明了在厌氧平移肌瘤中的OMP存在，因此牢固地增加了新兴视图，即Parenctomycetes具有革兰氏阴性细胞包络。

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