A Carrier Protein Strategy Yields the Structure of Dalbavancin
A Carrier Protein Strategy Yields the Structure of Dalbavancin
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DOI：
10.1021/ja208755j
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发表时间：
2012-03-14
影响因子：
15
通讯作者：
Loll, Patrick J.
中科院分区：
文献类型：
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作者：
Economou, Nicoleta J.;Nahoum, Virginie;Loll, Patrick J.
许多大的天然产物抗生素通过特异性结合和隔离在细菌细胞上发现的靶分子而起作用。我们已经开发了一种新的策略来加快这种抗生素靶复合物的结构分析，其中我们将靶分子共价连接到载体蛋白，然后使整个载体靶抗生素复合物结晶。使用天然化学连接，我们将糖肽抗生素的Lys-D-Ala-D-Ala结合表位连接到三种不同的载体蛋白。我们表明，多种抗生素对这种肽的识别不会因载体蛋白伴侣的存在而受到影响，并使用这种方法来确定新治疗药物达巴霉素的首次晶体结构。我们还报告了第一个晶体结构的不对称ristocycline抗生素二聚体，以及结构的万古霉素结合到载体靶融合。达巴万星的结构揭示了一种抗生素分子，它已经关闭了它的结合伴侣;它还表明了药物可以通过与血清蛋白结合来延长其半衰期的机制，并靶向细菌膜。值得注意的是，载体蛋白方法不限于肽配体如Lys-D-Ala-D-Ala，而是适用于多种靶标。这种策略可能会产生结构性的见解，加速新的治疗开发。
Many large natural product antibiotics act by specifically binding and sequestering target molecules found on bacterial cells. We have developed a new strategy to expedite the structural analysis of such antibiotic target complexes, in which we covalently link the target molecules to carrier proteins, and then crystallize the entire carrier target antibiotic complex. Using native chemical ligation, we have linked the Lys-D-Ala-D-Ala binding epitope for glycopeptide antibiotics to three different carrier proteins. We show that recognition of this peptide by multiple antibiotics is not compromised by the presence of the carrier protein partner, and use this approach to determine the first-ever crystal structure for the new therapeutic dalbavancin. We also report the first crystal structure of an asymmetric ristocetin antibiotic dimer, as well as the structure of vancomycin bound to a carrier target fusion. The dalbavancin structure reveals an antibiotic molecule that has closed around its binding partner; it also suggests mechanisms by which the drug can enhance its half-life by binding to serum proteins, and be targeted to bacterial membranes. Notably, the carrier protein approach is not limited to peptide ligands such as Lys-D-Ala-D-Ala, but is applicable to a diverse range of targets. This strategy is likely to yield structural insights that accelerate new therapeutic development.