Department of BioMedical Engineering, University of Groningen and University Medical Center Groningen, Groningen, Netherlands.
NPJ Biofilms Microbiomes. 2020 Aug 21;6(1):31. doi: 10.1038/s41522-020-00141-z.
Mechanosensitive channels in bacterial membranes open or close in response to environmental changes to allow transmembrane transport, including antibiotic uptake and solute efflux. In this paper, we hypothesize that gating of mechanosensitive channels is stimulated by forces through which bacteria adhere to surfaces. Hereto, channel gating is related with adhesion forces to different surfaces of a Staphylococcus aureus strain and its isogenic ΔmscL mutant, deficient in MscL (large) channel gating. Staphylococci becoming fluorescent due to uptake of calcein, increased with adhesion force and were higher in the parent strain (66% when adhering with an adhesion force above 4.0 nN) than in the ΔmscL mutant (40% above 1.2 nN). This suggests that MscL channels open at a higher critical adhesion force than at which physically different, MscS (small) channels open and contribute to transmembrane transport. Uptake of the antibiotic dihydrostreptomycin was monitored by staphylococcal killing. The parent strain exposed to dihydrostreptomycin yielded a CFU reduction of 2.3 log-units when adhering with an adhesion force above 3.5 nN, but CFU reduction remained low (1.0 log-unit) in the mutant, independent of adhesion force. This confirms that large channels open at a higher critical adhesion-force than small channels, as also concluded from calcein transmembrane transport. Collectively, these observations support our hypothesis that adhesion forces to surfaces play an important role, next to other established driving forces, in staphylococcal channel gating. This provides an interesting extension of our understanding of transmembrane antibiotic uptake and solute efflux in infectious staphylococcal biofilms in which bacteria experience adhesion forces from a wide variety of surfaces, like those of other bacteria, tissue cells, or implanted biomaterials.
细菌膜中的机械敏感通道会根据环境变化打开或关闭,以允许跨膜运输,包括抗生素摄取和溶质外排。在本文中,我们假设机械敏感通道的门控是由细菌附着在表面的力刺激的。为此,通道门控与金黄色葡萄球菌菌株及其同源 mscL 缺失突变体(mscL 通道门控缺陷)在不同表面的粘附力相关。由于摄取钙黄绿素,荧光金黄色葡萄球菌的数量随粘附力的增加而增加,并且在亲本菌株中(当粘附力大于 4.0nN 时为 66%)高于 mscL 缺失突变体(当粘附力大于 1.2nN 时为 40%)。这表明 MscL 通道在高于物理上不同的 MscS(小)通道打开并有助于跨膜运输的临界粘附力下打开。通过金黄色葡萄球菌的杀伤来监测抗生素二氢链霉素的摄取。当亲本菌株在粘附力大于 3.5nN 时暴露于二氢链霉素时,CFU 减少了 2.3 个对数单位,但在突变体中,CFU 减少仍然很低(1.0 个对数单位),与粘附力无关。这证实了大通道在高于小通道的临界粘附力下打开,这也与钙黄绿素跨膜转运的结论一致。总之,这些观察结果支持我们的假设,即除了其他已建立的驱动力外,表面的粘附力在金黄色葡萄球菌通道门控中也起着重要作用。这为我们理解感染性金黄色葡萄球菌生物膜中跨膜抗生素摄取和溶质外排提供了一个有趣的扩展,在生物膜中,细菌会经历来自各种表面的粘附力,如其他细菌、组织细胞或植入的生物材料。