Interaction between cell-penetrating peptides and acid-sensitive anionic oligopeptides as a model for the design of targeted drug carriers.
Interaction between cell-penetrating peptides and acid-sensitive anionic oligopeptides as a model for the design of targeted drug carriers.
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DOI：
10.1021/mp400747k
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发表时间：
2014-05-05
影响因子：
4.9
通讯作者：
Zaro JL
中科院分区：
文献类型：
--
作者：
Sun C;Shen WC;Tu J;Zaro JL
克服细胞穿透肽(CPPs)的非特异性细胞摄取是其临床应用的主要障碍。以pH为激活开关，将组氨酸-谷氨酸(HE)二肽重复序列与CPPs融合，在pH为6.5或以下的中等酸性环境中激发穿膜活性，在中性pH(即pH 7.0或以上)下屏蔽内化作用。在本研究中，设计了一系列含有HE寡肽序列(即(HE)n，n=8、10或12)和阳离子CPP(即YG(RG)6、YGR6G6或TAT)的重组GST融合蛋白，用于pH敏感研究，比较它们在培养的HeLa细胞中的摄取和表面结合。用圆二色谱(CD)分析了CPP的二级结构差异与pH敏感性之间的关系。具有聚集精氨酸残基的YGR6G6比具有分离精氨酸残基的YG(RG)6对细胞摄取的pH更敏感。增加HE重复序列的延伸降低了YG(RG)6和YGR6G6的细胞摄取和表面结合。细胞内在化比率在pH 7.5vs6.0时不受血清的影响。CD光谱分析表明，(HE)10-TAT和(HE)10-YGR6G6均为无序二级结构，而(HE)10-YG(RG)6为反平行β-Sheet构象。这种β-Sheet构象可能稳定了(HE)10与YG(RG)6的结合，导致(HE)10-YG(RG)6的pH敏感性减弱。另一方面，细胞实验表明，(HE)10-YGR6G6和(HE)10-TAT的无规卷曲结构都表现出更高的pH敏感性。本研究为将来设计pH敏感的HE-CPP靶向给药载体奠定了基础。
Overcoming the nonspecific cellular uptake of cell-penetrating peptides (CPPs) is a major hurdle in their clinical application. Using pH as the activation switch, histidine-glutamic acid (HE) dipeptide repeats were fused to CPPs to trigger the membrane-penetrating activity at mildly acidic pH environments (i.e., pH 6.5 or below) while masking the internalization at neutral pH (i.e., pH 7.0 or above). In this study, a series of recombinant GST-fusion proteins containing an HE oligopeptide sequence (i.e., (HE)n with n = 8, 10, or 12) and a cationic CPP (i.e., YG(RG)6, YGR6G6, or Tat) were engineered for a pH-sensitive study comparing their cellular uptake and surface binding in cultured HeLa cells. Circular dichroism (CD) spectroscopy was performed to correlate differences between CPPs in secondary structure with the pH sensitivity. YGR6G6 with clustered arginine residues exhibited greater pH sensitivity in cellular uptake than YG(RG)6 with separated arginine residues. Increasing the stretch of HE repeats decreased cellular uptake and surface binding for both YG(RG)6 and YGR6G6. The ratio of cellular internalization at pH 7.5 vs 6.0 was not changed by the presence of serum. CD spectral data revealed that both (HE)10-Tat and (HE)10-YGR6G6 exhibited an unordered secondary structure, whereas (HE)10-YG(RG)6 adopted an antiparallel β-sheet conformation. This β-sheet conformation presumably stabilized the association of (HE)10 with YG(RG)6, leading to weakened pH sensitivity of (HE)10-YG(RG)6. On the other hand, the random-coiled structures, that is, (HE)10-YGR6G6 and (HE)10-Tat, both showed higher pH sensitivity as determined in cell experiments. The data presented in this study provide a basis for the future design of pH-sensitive HE-CPP carrier for targeted drug delivery.
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影响因子：
14
作者：
Fei, Likun;Yap, Li-Peng;Conti, Peter S.;Shen, Wei-Chiang;Zaro, Jennica L.
通讯作者：
Zaro, Jennica L.
影响因子：
14
作者：
Liu, Betty R.;Huang, Yue-wern;Lee, Han-Jung
通讯作者：
Lee, Han-Jung
影响因子：
14.8
作者：
Greenfield, Norma J.
通讯作者：
Greenfield, Norma J.
利用细胞穿透肽的力量：可激活载体，用于靶向全身递送癌症治疗剂和显像剂。
DOI：
10.1002/wnan.1197
发表时间：
2013-01
影响因子：
8.6
作者：
MacEwan, Sarah R.;Chilkoti, Ashutosh
通讯作者：
Chilkoti, Ashutosh
影响因子：
4.8
作者：
Eguchi, A;Akuta, T;Nakanishi, M
通讯作者：
Nakanishi, M