A new survivin tracer tracks, delocalizes and captures endogenous survivin at different subcellular locations and in distinct organelles.
A new survivin tracer tracks, delocalizes and captures endogenous survivin at different subcellular locations and in distinct organelles.
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DOI：
10.1038/srep31177
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发表时间：
2016-08-12
影响因子：
4.6
通讯作者：
Gettemans J
中科院分区：
文献类型：
--
作者：
Beghein E;Van Audenhove I;Zwaenepoel O;Verhelle A;De Ganck A;Gettemans J
Survivin是凋亡抑制蛋白家族中最小的成员，在细胞有丝分裂过程中起着重要作用，具有细胞保护功能。存活素在大多数癌症类型中高度表达，并有助于癌症发生的多个方面。其高度多样化的功能背后的分子机制需要广泛探索，这对于未来个性化治疗的合理设计至关重要。在这项研究中，我们已经产生了一种羊驼生存素纳米抗体（SVVNb8），它以低纳摩尔亲和力与其靶标结合。当在HeLa细胞中表达为胞内抗体时，SVVNb8在有丝分裂的不同阶段忠实地跟踪生存素而不干扰生存素功能。此外，偶联SVVNb8与亚细胞离域标签有效地将内源性存活素重定向到细胞核、细胞质、过氧化物酶体，甚至到线粒体的膜间隙，在那里它可能与驻留的线粒体存活素相互作用。基于我们的研究结果，我们认为SVVNb8是一个很好的工具，以进一步阐明生存素的生物学和地形，并可以作为一个模型系统，研究线粒体和过氧化物酶体（生存素）蛋白的进口。
Survivin, the smallest member of the inhibitor of apoptosis protein family, plays a central role during mitosis and exerts a cytoprotective function. Survivin is highly expressed in most cancer types and contributes to multiple facets of carcinogenesis. The molecular mechanisms underlying its highly diverse functions need to be extensively explored, which is crucial for rational design of future personalized therapeutics. In this study, we have generated an alpaca survivin nanobody (SVVNb8) that binds with low nanomolar affinity to its target. When expressed as an intrabody in HeLa cells, SVVNb8 faithfully tracks survivin during different phases of mitosis without interfering with survivin function. Furthermore, coupling SVVNb8 with a subcellular delocalization tag efficiently redirects endogenous survivin towards the nucleus, the cytoplasm, peroxisomes and even to the intermembrane space of mitochondria where it presumably interacts with resident mitochondrial survivin. Based on our findings, we believe that SVVNb8 is an excellent instrument to further elucidate survivin biology and topography, and can serve as a model system to investigate mitochondrial and peroxisomal (survivin) protein import.