A structural basis for cellular uptake of GST-fold proteins.
A structural basis for cellular uptake of GST-fold proteins.
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DOI：
10.1371/journal.pone.0017864
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发表时间：
2011-03-24
期刊：
影响因子：
3.7
通讯作者：
Casarotto MG
中科院分区：
文献类型：
--
作者：
Morris MJ;Liu D;Weaver LM;Board PG;Casarotto MG
最近发现，谷胱甘肽转移酶（GST）和其他结构相关分子可以从外部介质转移到许多不同的细胞类型中。在这项研究中，我们旨在详细探索控制细胞易位的结构特征，并通过解剖人类 GST 酶 GSTM2-2，我们定量证明 α-螺旋 C 端结构域 (GST-C) 负责这种特性。尝试进一步检查 GST-C 内的组成螺旋导致细胞易位效率降低，表明内在的 GST-C 结构域结构对于最大细胞易位能力是必要的。特别值得注意的是，GST-C 的 α-6 螺旋在此结构域的折叠中发挥稳定作用。通过破坏 GST-C 构象的稳定性，观察到细胞易位效率提高了约 2 倍。通过圆二色性和差示扫描荧光测量研究了这些蛋白质构建体的结构稳定性概况，并发现其对其细胞易位效率有影响。这些实验表明，“GST-折叠”结构基序中的球状螺旋结构域在影响细胞摄取方面发挥着作用，而影响 GST-C 构象稳定性的变化可以显着影响细胞转位效率。
It has recently emerged that glutathione transferase enzymes (GSTs) and other structurally related molecules can be translocated from the external medium into many different cell types. In this study we aim to explore in detail, the structural features that govern cell translocation and by dissecting the human GST enzyme GSTM2-2 we quantatively demonstrate that the α-helical C-terminal domain (GST-C) is responsible for this property. Attempts to further examine the constituent helices within GST-C resulted in a reduction in cell translocation efficiency, indicating that the intrinsic GST-C domain structure is necessary for maximal cell translocation capacity. In particular, it was noted that the α-6 helix of GST-C plays a stabilising role in the fold of this domain. By destabilising the conformation of GST-C, an increase in cell translocation efficiency of up to ∼2-fold was observed. The structural stability profiles of these protein constructs have been investigated by circular dichroism and differential scanning fluorimetry measurements and found to impact upon their cell translocation efficiency. These experiments suggest that the globular, helical domain in the ‘GST-fold’ structural motif plays a role in influencing cellular uptake, and that changes that affect the conformational stability of GST-C can significantly influence cell translocation efficiency.
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影响因子：
3.4
作者：
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通讯作者：
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