Inhibition of the β-barrel assembly machine by a peptide that binds BamD
Inhibition of the β-barrel assembly machine by a peptide that binds BamD
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DOI：
10.1073/pnas.1415955112
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发表时间：
2015-02-17
影响因子：
11.1
通讯作者：
Kahne, Daniel
中科院分区：
文献类型：
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作者：
Hagan, Christine L.;Wzorek, Joseph S.;Kahne, Daniel
在革兰氏阴性细菌的外膜中组装积分膜β-桶蛋白的蛋白质复合物是开发新抗生素的一个有吸引力的靶标。该复合物是β-桶装配机（BAM），其中包含两个必需蛋白质，Bama和Bamd。我们已经确定了一种肽，该肽通过表征BAMD与展开的底物蛋白的相互作用，从而在体外抑制β-桶蛋白的组装。该肽是底物蛋白的片段，并包含一个保守的氨基酸序列。我们已经证明，在全长底物蛋白质中，该序列的突变损害了蛋白质的组装，这表明BAMD与该序列的相互作用是组装机制的重要组成部分。最后，我们发现含有该序列的肽的体内表达会导致生长缺陷，并使大肠杆菌对通常具有抗性的抗生素敏感。因此，抑制底物与BAMD的结合是开发针对革兰氏阴性细菌的新抗生素的可行策略。
The protein complex that assembles integral membrane beta-barrel proteins in the outer membranes of Gram-negative bacteria is an attractive target in the development of new antibiotics. This complex, the beta-barrel assembly machine (Bam), contains two essential proteins, BamA and BamD. We have identified a peptide that inhibits the assembly of beta-barrel proteins in vitro by characterizing the interaction of BamD with an unfolded substrate protein. This peptide is a fragment of the substrate protein and contains a conserved amino acid sequence. We have demonstrated that mutations of this sequence in the full-length substrate protein impair the protein's assembly, implying that BamD's interaction with this sequence is an important part of the assembly mechanism. Finally, we have found that in vivo expression of a peptide containing this sequence causes growth defects and sensitizes Escherichia coli to antibiotics to which they are normally resistant. Therefore, inhibiting the binding of substrates to BamD is a viable strategy for developing new antibiotics directed against Gram-negative bacteria.