Willems Kherim, Van Meervelt Veerle, Wloka Carsten, Maglia Giovanni
Department of Chemistry, KU Leuven, Celestijnenlaan 200G, 3001 Leuven, Belgium.
Department of Life Sciences and Imaging, IMEC, Kapeldreef 75, 3001 Leuven, Belgium.
Philos Trans R Soc Lond B Biol Sci. 2017 Aug 5;372(1726). doi: 10.1098/rstb.2016.0230.
Biological nanopores are a class of membrane proteins that open nanoscale water conduits in biological membranes. When they are reconstituted in artificial membranes and a bias voltage is applied across the membrane, the ionic current passing through individual nanopores can be used to monitor chemical reactions, to recognize individual molecules and, of most interest, to sequence DNA. In addition, a more recent nanopore application is the analysis of single proteins and enzymes. Monitoring enzymatic reactions with nanopores, i.e. nanopore enzymology, has the unique advantage that it allows long-timescale observations of native proteins at the single-molecule level. Here, we describe the approaches and challenges in nanopore enzymology.This article is part of the themed issue 'Membrane pores: from structure and assembly, to medicine and technology'.
生物纳米孔是一类能在生物膜中打开纳米级水通道的膜蛋白。当它们被重组到人工膜中并在膜上施加偏置电压时，通过单个纳米孔的离子电流可用于监测化学反应、识别单个分子，最受关注的是用于DNA测序。此外，纳米孔的一个最新应用是对单个蛋白质和酶进行分析。用纳米孔监测酶促反应，即纳米孔酶学，具有独特的优势，它能在单分子水平上对天然蛋白质进行长时间尺度的观察。在此，我们描述纳米孔酶学中的方法和挑战。本文是主题为“膜孔：从结构与组装到医学与技术”这一特刊的一部分。
