Katz B Z, Krylov D, Aota S, Olive M, Vinson C, Yamada K M
National Institute of Dental Research, NIH, Bethesda, MD, USA.
Biotechniques. 1998 Aug;25(2):298-302, 304. doi: 10.2144/98252rr01.
Green fluorescent protein (GFP) is a valuable marker for intracellular protein localization. However the fusion of GFP with structural proteins can alter their properties, resulting in a loss of fusion protein localization, decreased GFP fluorescence or both. We describe a novel targeting approach based on noncovalent heterodimerization of GFP and cytoplasmic structural proteins. The formation of structural protein/GFP complexes was mediated by modified leucine zipper protein spacers designed to form high-affinity heterodimers. The complexes localized accurately to specific sites within cells, providing selective fluorescence labeling of subcellular structures such as microfilaments or focal contacts.
绿色荧光蛋白（GFP）是用于细胞内蛋白质定位的一种重要标记物。然而，GFP与结构蛋白的融合可能会改变它们的特性，导致融合蛋白定位丧失、GFP荧光减弱或两者皆有。我们描述了一种基于GFP与细胞质结构蛋白非共价异源二聚化的新型靶向方法。结构蛋白/GFP复合物的形成由经过修饰的亮氨酸拉链蛋白间隔区介导，该间隔区设计用于形成高亲和力异源二聚体。这些复合物能够准确地定位于细胞内的特定部位，为亚细胞结构如微丝或黏着斑提供选择性荧光标记。
