Tat peptide is capable of importing large nanoparticles across nuclear membrane in digitonin permeabilized cells.
Tat peptide is capable of importing large nanoparticles across nuclear membrane in digitonin permeabilized cells.
复制标题
DOI：
10.1007/s10439-009-9768-0
复制
发表时间：
2009-10
影响因子：
3.8
通讯作者：
Bao, Gang
中科院分区：
文献类型：
--
作者：
Nitin, Nitin;LaConte, Leslie;Rhee, Won Jong;Bao, Gang
了解核进口的能力和局限性对于有效地运送用于疾病诊断和靶向治疗的大分子和纳米颗粒至关重要。在这里，我们报道了TAT多肽介导的不同的货物进入细胞核，包括染料标记的链霉亲和素蛋白，43和90 nm荧光小球，以及用于动力学测量的∼20 nm量子点。我们的结果揭示了TAT和NLS介导的核进口之间的显著差异：与NLS递送不同，基于TAT多肽的递送不受WGA阻断的抑制，也不需要ATP。令人惊讶的是，TAT多肽能够将90 nm的小球导入洋地黄素通透性细胞的细胞核，这表明其与核膜的相互作用遵循不同于NLS的机制。用TAT多肽标记的量子点定量了导入动力学，得到动力学常数为0.0085 S−1。综上所述，我们的结果表明，与NLS相比，TAT多肽介导的核导入更快，遵循不同的途径，并且能够导入大尺寸纳米颗粒。这些结果对开发将货物运送到活细胞核中的新方法具有重要意义。
Understanding the capabilities and limitations of nuclear import is crucial to efficient delivery of macromolecules and nanoparticles for diagnosis and targeted therapy of diseases. Here we report the Tat peptide-mediated import of different cargos into cell nucleus, including dye-labeled streptavidin protein, 43 and 90 nm fluorescent beads, as well as ∼20 nm quantum dots for kinetic measurements. Our results revealed significant differences between Tat- and NLS-mediated nuclear import: unlike delivery with the NLS, Tat peptide-based delivery is not inhibited by WGA blockage nor does it require ATP. Surprisingly, Tat peptide was able to import 90 nm beads into the nuclei of digitonin-permeabilized cells, suggesting that its interaction with the nuclear envelope follows a mechanism different from that of NLS. The import kinetics was quantified using Tat peptide-conjugated QDs, yielding a kinetic constant of 0.0085 s−1. Taken together, our results suggest that, compared with NLS, Tat peptide-mediated nuclear import is faster, follows a different pathway, and is capable of importing large nanoparticles. These results have significant implications for the development of new approaches for delivery of cargo into the nuclei of living cells.
登录
查看更多内容
影响因子：
11.4
作者：
Ribbeck, K;Görlich, D
通讯作者：
Görlich, D
影响因子：
4.8
作者：
Wilson, GL;Dean, BS;Dean, DA
通讯作者：
Dean, DA
影响因子：
3.3
作者：
Panté, N;Kann, M
通讯作者：
Kann, M
影响因子：
4.8
作者：
Becker-Hapak, M;McAllister, SS;Dowdy, SF
通讯作者：
Dowdy, SF
核蛋白在透化的哺乳动物细胞中进口需要可溶性细胞质因子。
影响因子：
7.8
作者：
Adam, S A;Marr, R S;Gerace, L
通讯作者：
Gerace, L