Petrache A Ivona, Machin Darren C, Williamson Daniel J, Webb Michael E, Beales Paul A
School of Chemistry and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK.
Mol Biosyst. 2016 May 24;12(6):1760-3. doi: 10.1039/c6mb00126b.
Lipid nanodiscs have broad applications in membrane protein assays, biotechnology and materials science. Chemical modification of the nanodiscs to expand their functional attributes is generally desirable for all of these uses. We present a method for site-selective labelling of the N-terminus of the nanodisc's membrane scaffold protein (MSP) using the Sortase A protein. Labelling of the MSP was achieved when assembled within the lipid nanodisc architecture, demonstrating that this method can be used as a retrofit approach to modification of preformed nanodiscs before or during application. We label the MSP with a fluorescent fluorescein moiety and use them to image nanodisc uptake into HeLa cells. The Sortase A labelling method could be employed as a general approach to labelling nanodiscs with application-specific functionalities.
脂质纳米盘在膜蛋白分析、生物技术和材料科学领域有着广泛的应用。对纳米盘进行化学修饰以扩展其功能属性，对于所有这些应用来说通常都是很有必要的。我们提出了一种使用分选酶A蛋白对纳米盘的膜支架蛋白（MSP）的N端进行位点选择性标记的方法。当MSP在脂质纳米盘结构内组装时实现了标记，这表明该方法可作为一种在应用前或应用过程中对预制纳米盘进行修饰的改造方法。我们用荧光素部分对MSP进行标记，并利用它们来成像纳米盘被HeLa细胞摄取的情况。分选酶A标记方法可作为一种用特定应用功能标记纳米盘的通用方法。
