Constructing fluorogenic Bacillus spores (F-spores) via hydrophobic decoration of coat proteins.
Constructing fluorogenic Bacillus spores (F-spores) via hydrophobic decoration of coat proteins.
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DOI：
10.1371/journal.pone.0009283
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发表时间：
2010-02-19
期刊：
影响因子：
3.7
通讯作者：
Rotman B
中科院分区：
文献类型：
--
作者：
Ferencko L;Rotman B
细菌孢子由大约60种不同的蛋白质组成的涂层保护，这些蛋白质组装成具有有趣的形态和机械性能的生物化学复杂结构。历史上，外套被认为是一种静态结构，提供刚性，主要作为筛子来排除外部大的有毒分子，如裂解酶。然而，近年来，通过使用自动扫描显微镜和高分辨率原子力显微镜等创新工具进行的实验，出现了关于涂层结构和功能的新信息。使用薄片电子显微镜，我们发现芽孢杆菌孢子的外壳具有拓扑特异性的蛋白质，形成了一层可识别的层，因为它自发地被来自环境的疏水荧光探针装饰。此外，带有修饰外壳蛋白的孢子(称为F孢子)具有通过产生强烈的荧光来响应外部萌发信号的意想不到的属性。来自不同实验设计的荧光数据，包括来自五个不同芽孢杆菌的F-孢子，表明F-孢子的荧光能力受假定的萌发依赖机制的控制。这项工作揭示了孢子壳蛋白的一个新属性，我们利用它来修饰特定的一层，赋予F-孢子依赖于萌发的荧光原性。我们期望F-孢子将提供一个模型系统，以获得对孢子壳蛋白结构/功能动态的新见解。
Bacterial spores are protected by a coat consisting of about 60 different proteins assembled as a biochemically complex structure with intriguing morphological and mechanical properties. Historically, the coat has been considered a static structure providing rigidity and mainly acting as a sieve to exclude exogenous large toxic molecules, such as lytic enzymes. Over recent years, however, new information about the coat's architecture and function have emerged from experiments using innovative tools such as automated scanning microscopy, and high resolution atomic force microscopy. Using thin-section electron microscopy, we found that the coat of Bacillus spores has topologically specific proteins forming a layer that is identifiable because it spontaneously becomes decorated with hydrophobic fluorogenic probes from the milieu. Moreover, spores with decorated coat proteins (termed F-spores) have the unexpected attribute of responding to external germination signals by generating intense fluorescence. Fluorescence data from diverse experimental designs, including F-spores constructed from five different Bacilli species, indicated that the fluorogenic ability of F-spores is under control of a putative germination-dependent mechanism. This work uncovers a novel attribute of spore-coat proteins that we exploited to decorate a specific layer imparting germination-dependent fluorogenicity to F-spores. We expect that F-spores will provide a model system to gain new insights into structure/function dynamics of spore-coat proteins.
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DOI：
10.1016/j.bbrc.2004.05.062
发表时间：
2004-07-02
影响因子：
3.1
作者：
Ferencko, L;Cote, MA;Rotman, B
通讯作者：
Rotman, B
影响因子：
3.2
作者：
ROBINOW, CF
通讯作者：
ROBINOW, CF
影响因子：
6.8
作者：
Goh, CS;Milburn, D;Gerstein, M
通讯作者：
Gerstein, M
影响因子：
3.2
作者：
Paidhungat, M;Ragkousi, K;Setlow, P
通讯作者：
Setlow, P
影响因子：
4
作者：
Lavis, Luke D.
通讯作者：
Lavis, Luke D.