Arnoys Eric J, Ackerman Cheri M, Wang John L
Department of Chemistry and Biochemistry, Calvin College, Grand Rapids, MI, USA.
Methods Mol Biol. 2015;1207:465-83. doi: 10.1007/978-1-4939-1396-1_30.
A large number of observations on the nuclear versus cytoplasmic distribution of galectin-3 have been reported, correlating the presence or absence of the protein in a particular compartment of the cell to various parameters such as source of the cells under study, specific cell type, culture conditions, proliferation status of the cell/culture, or neoplastic transformation. In fact, galectin-3 exhibits the phenomenon of nucleocytoplasmic shuttling, defined as the repeated bidirectional movement of a protein across the nuclear pore complex. Nevertheless, the finding that galectin-3 can show a predominantly nuclear localization under one set of conditions and a prominent cytoplasmic localization under other conditions suggests specific and regulated mechanisms of balance between cytoplasmic anchorage, nuclear import, nuclear retention, and nuclear export. One key consideration in the understanding of these processes is the definition of the signals and receptors that mediate the transport. In this chapter, we describe the experimental procedures that have allowed us to document the phenomenon of nucleocytoplasmic shuttling and the identification of the nuclear localization signal as well as the nuclear export signal.
关于半乳糖凝集素-3在细胞核与细胞质中的分布,已有大量观察报道,将该蛋白在细胞特定区室中的存在与否与各种参数相关联,如所研究细胞的来源、特定细胞类型、培养条件、细胞/培养物的增殖状态或肿瘤转化。事实上,半乳糖凝集素-3表现出核质穿梭现象,即蛋白质反复双向穿过核孔复合体的运动。然而,半乳糖凝集素-3在一组条件下可主要定位于细胞核,而在其他条件下则主要定位于细胞质,这一发现表明在细胞质锚定、核输入、核滞留和核输出之间存在特定且受调控的平衡机制。理解这些过程的一个关键因素是介导运输的信号和受体的定义。在本章中,我们描述了一些实验程序,这些程序使我们能够记录核质穿梭现象,并鉴定核定位信号以及核输出信号。