Periago Jessica, Mason Clarissa, Griep Mark A
Department of Chemistry, University of Nebraska-Lincoln, Lincoln, Nebraska 68588-0304, United States.
ACS Omega. 2022 Mar 1;7(10):8420-8428. doi: 10.1021/acsomega.1c05928. eCollection 2022 Mar 15.
The widespread use of antibiotics to treat infections is one of the reasons that global mortality rates have fallen over the past 80 years. However, antibiotic use is also responsible for the concomitant rise in antibiotic resistance because it results in dysbiosis in which commensal and pathogenic bacteria are both greatly reduced. Therefore, narrow-range antibiotics are a promising direction for reducing antibiotic resistance because they are more discriminate. As a step toward addressing this problem, the goal of this study was to identify sites on DnaG primase that are conserved within Gram-positive bacteria and different from the equivalent sites in Gram-negative bacteria. Based on sequence and structural analysis, the primase C-terminal helicase-binding domain (CTD) was identified as most promising. Although the primase CTD sequences are very poorly conserved, they have highly conserved protein folds, and Gram-positive bacterial primases fold into a compact state that creates a small molecule binding site adjacent to a groove. The small molecule would stabilize the protein in its compact state, which would interfere with the helicase binding. This is important because primase CTD must be in its open conformation to bind to its cognate helicase at the replication fork.
使用抗生素治疗感染的广泛应用是过去80年全球死亡率下降的原因之一。然而,抗生素的使用也导致了抗生素耐药性的相应增加,因为它会导致生态失调,共生菌和病原菌都会大幅减少。因此,窄谱抗生素是降低抗生素耐药性的一个有前景的方向,因为它们更具选择性。作为解决这一问题的一步,本研究的目标是确定DnaG引发酶上在革兰氏阳性菌中保守且不同于革兰氏阴性菌中对应位点的位点。基于序列和结构分析,引发酶C端解旋酶结合结构域(CTD)被确定为最有前景的。尽管引发酶CTD序列的保守性很差,但它们具有高度保守的蛋白质折叠,革兰氏阳性菌引发酶折叠成紧凑状态,形成一个与凹槽相邻的小分子结合位点。小分子会使蛋白质稳定在其紧凑状态,这会干扰解旋酶的结合。这很重要,因为引发酶CTD必须处于开放构象才能在复制叉处与同源解旋酶结合。