Export of Diverse and Bioactive Small Proteins through a Type I Secretion System.
Export of Diverse and Bioactive Small Proteins through a Type I Secretion System.
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DOI：
10.1128/aem.00335-23
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发表时间：
2023-05-31
影响因子：
4.4
通讯作者：
中科院分区：
文献类型：
--
作者：
小蛋白质执行各种各样的功能，从微生物竞争到内分泌信号，再到构建生物材料。可以产生重组小蛋白的微生物系统使得能够发现新的效应物、探索序列活性关系，并且具有体内递送的潜力。然而，我们缺乏简单的系统来控制革兰氏阴性菌的小蛋白分泌。微菌素是由革兰氏阴性菌分泌的小蛋白抗生素，可抑制邻近微生物的生长。它们通过一类特定的I型分泌系统（T1SSs）以一步过程从胞质溶胶输出到环境中。然而，相对较少的是已知的小蛋白质出口通过microcin T1SS的底物要求。在这里，我们研究了原型microcin V T1SS从大肠杆菌，并表明，它可以输出一个非常广泛的天然和合成的小蛋白。我们表明，分泌在很大程度上是独立的货物蛋白的化学性质，似乎只受蛋白质长度。我们表明，各种各样的生物活性序列，包括抗菌蛋白，微生物信号传导因子，蛋白酶抑制剂，和人类激素，都可以分泌，并引发其预期的生物效应。通过该系统的分泌并不限于E.大肠杆菌，我们证明了它的功能，在其他革兰氏阴性菌，可以栖息在胃肠道。我们的研究结果揭示了小蛋白通过微菌素V T1SS出口的高度混杂性，这对天然货物容量以及该系统在革兰氏阴性细菌中用于小蛋白研究和交付具有影响。革兰氏阴性菌中分泌微粒蛋白的I型分泌系统在一个步骤中将小的抗菌蛋白从细胞质转运到细胞外环境。在自然界中，每个分泌系统通常与特定的小蛋白配对。我们对这些转运蛋白的输出能力以及货物序列如何影响分泌知之甚少。在这里，我们调查的microcin V I型系统。值得注意的是，我们的研究表明，该系统可以输出不同序列组成的小蛋白质，并且仅受蛋白质长度的限制。此外，我们证明了广泛的生物活性小蛋白可以分泌，该系统可用于革兰氏阴性菌种，殖民胃肠道。这些发现扩展了我们对I型系统分泌的理解，以及它们在各种小蛋白应用中的潜在用途。
Small proteins perform a diverse array of functions, from microbial competition, to endocrine signaling, to building biomaterials. Microbial systems that can produce recombinant small proteins enable discovery of new effectors, exploration of sequence activity relationships, and have the potential for in vivo delivery. However, we lack simple systems for controlling small-protein secretion from Gram-negative bacteria. Microcins are small-protein antibiotics secreted by Gram-negative bacteria that inhibit the growth of neighboring microbes. They are exported from the cytosol to the environment in a one-step process through a specific class of type I secretion systems (T1SSs). However, relatively little is known about substrate requirements for small proteins exported through microcin T1SSs. Here, we investigate the prototypic microcin V T1SS from Escherichia coli and show that it can export a remarkably wide range of natural and synthetic small proteins. We demonstrate that secretion is largely independent of the cargo protein’s chemical properties and appears to be constrained only by protein length. We show that a varied range of bioactive sequences, including an antibacterial protein, a microbial signaling factor, a protease inhibitor, and a human hormone, can all be secreted and elicit their intended biological effect. Secretion through this system is not limited to E. coli, and we demonstrate its function in additional Gram-negative species that can inhabit the gastrointestinal tract. Our findings uncover the highly promiscuous nature of small-protein export through the microcin V T1SS, which has implications for native-cargo capacity and the use of this system in Gram-negative bacteria for small-protein research and delivery. IMPORTANCE Type I secretion systems for microcin export in Gram-negative bacteria transport small antibacterial proteins from the cytoplasm to the extracellular environment in a single step. In nature, each secretion system is generally paired with a specific small protein. We know little about the export capacity of these transporters and how cargo sequence influences secretion. Here, we investigate the microcin V type I system. Remarkably, our studies show that this system can export small proteins of diverse sequence composition and is only limited by protein length. Furthermore, we demonstrate that a wide range of bioactive small proteins can be secreted and that this system can be used in Gram-negative species that colonize the gastrointestinal tract. These findings expand our understanding of secretion through type I systems and their potential uses in a variety of small-protein applications.
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影响因子：
3.7
作者：
Green ER;Mecsas J
通讯作者：
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