Månsson Alf, Sundberg Mark, Balaz Martina, Bunk Richard, Nicholls Ian A, Omling Pär, Tågerud Sven, Montelius Lars
Department of Chemistry and Biomedical Sciences, University of Kalmar, SE-39182 Kalmar, Sweden.
Biochem Biophys Res Commun. 2004 Feb 6;314(2):529-34. doi: 10.1016/j.bbrc.2003.12.133.
We recently refined the in vitro motility assay for studies of actomyosin function to achieve rectified myosin induced sliding of actin filaments. This paves the way, both for detailed functional studies of actomyosin and for nanotechnological applications. In the latter applications it would be desirable to use actin filaments for transportation of cargoes (e.g., enzymes) between different predetermined locations on a chip. We here describe how single quantum dot labelling of isolated actin filaments simultaneously provides handles for cargo attachment and bright and photostable fluorescence labels facilitating cargo detection and filament tracking. Labelling was achieved with preserved actomyosin function using streptavidin-coated CdSe quantum dots (Qdots). These nanocrystals have several unique physical properties and the present work describes their first use for functional studies of isolated proteins outside the cell. The results, in addition to the nanotechnology developments, open for new types of in vitro assays of isolated biomolecules.
我们最近改进了用于研究肌动球蛋白功能的体外运动分析方法，以实现肌球蛋白诱导的肌动蛋白丝的整流滑动。这为肌动球蛋白的详细功能研究和纳米技术应用都铺平了道路。在后者的应用中，希望使用肌动蛋白丝在芯片上不同的预定位置之间运输货物（例如酶）。我们在此描述了如何对分离的肌动蛋白丝进行单量子点标记，同时为货物附着提供手柄，并提供明亮且光稳定的荧光标记，便于货物检测和丝跟踪。使用链霉亲和素包被的CdSe量子点（Qdots）实现了在保留肌动球蛋白功能的情况下进行标记。这些纳米晶体具有几种独特的物理特性，并且本工作描述了它们首次用于细胞外分离蛋白质的功能研究。除了纳米技术的发展之外，这些结果还为新型的体外分离生物分子分析方法开辟了道路。
