Van Veen Sarah, Martin Shaun, Schuermans Marleen, Vangheluwe Peter
Department of Cellular and Molecular Medicine, KU Leuven, Leuven, Belgium.
Bio Protoc. 2021 Jan 20;11(2):e3888. doi: 10.21769/BioProtoc.3888.
ATP13A2/PARK9 is a late endo-/lysosomal P5B transport ATPase that is associated with several neurodegenerative disorders. We recently characterized ATP13A2 as a lysosomal polyamine exporter, which sheds light on the molecular identity of the unknown mammalian polyamine transport system. Here, we describe step by step a protocol to measure radiolabeled polyamine transport in reconstituted vesicles from yeast cells overexpressing human ATP13A2. This protocol was developed as part of our recent publication (van Veen , 2020 ) and will be useful for characterizing the transport function of other putative polyamine transporters, such as isoforms of the P5B transport ATPases.
ATP13A2/PARK9是一种晚期内体/溶酶体P5B转运ATP酶，与多种神经退行性疾病相关。我们最近将ATP13A2鉴定为溶酶体多胺输出蛋白，这为未知的哺乳动物多胺转运系统的分子特性提供了线索。在这里，我们逐步描述了一种用于测量过表达人ATP13A2的酵母细胞重构囊泡中放射性标记多胺转运的实验方案。该方案是我们最近发表的论文（van Veen，2020）的一部分，对于表征其他假定的多胺转运蛋白（如P5B转运ATP酶的同工型）的转运功能将是有用的。
