Tailoring Cationic Helical Polypeptides for Efficient Cytosolic Protein Delivery

doi:10.1007/s40242-019-0060-z

高等学校化学研究 ›› 2020, Vol. 36 ›› Issue (1): 134-138.doi: 10.1007/s40242-019-0060-z

Tailoring Cationic Helical Polypeptides for Efficient Cytosolic Protein Delivery

WANG Ruijue, SHENG Kai, HOU Yingqin, SUN Jialing, LU Hua

Beijing National Laboratory for Molecular Sciences, Center for Soft Matter Science and Engineering, Key Laboratory of Polymer Chemistry and Physics of Ministry of Education, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, P. R. China

收稿日期:2019-11-22 修回日期:2019-12-18 出版日期:2020-02-01 发布日期:2019-12-18
通讯作者: LU Hua E-mail:chemhualu@pku.edu.cn
基金资助:
Supported by the National Key Research and Development Program of China(No.2016YFA0201400) and the National Natural Science Foundation of China(Nos.21434008, 21722401).

Tailoring Cationic Helical Polypeptides for Efficient Cytosolic Protein Delivery

WANG Ruijue, SHENG Kai, HOU Yingqin, SUN Jialing, LU Hua

Beijing National Laboratory for Molecular Sciences, Center for Soft Matter Science and Engineering, Key Laboratory of Polymer Chemistry and Physics of Ministry of Education, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, P. R. China

Received:2019-11-22 Revised:2019-12-18 Online:2020-02-01 Published:2019-12-18
Contact: LU Hua E-mail:chemhualu@pku.edu.cn
Supported by:
Supported by the National Key Research and Development Program of China(No.2016YFA0201400) and the National Natural Science Foundation of China(Nos.21434008, 21722401).

摘要/Abstract

摘要： Protein delivery is of central importance for both diagnostic and therapeutic applications. However, protein delivery faces challenges including poor endosomal escape and thus limited efficiency. Here, we report the facile construction and screening of a small library of cationic helical polypeptides for cytosolic protein delivery. The library is based on a random copolymer poly(γ-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}esteryl-L-glutamate)-randompoly(γ-6-chlorohexyl-L-glutamate)[P(EG₃-r-ClC₆)Glu], which is then modified with various pyridine derivatives and alkyl thiols. Flow Cytometry, confocal laser scanning microscopy, and viability assay collaboratively identify two leading polymers, showing efficient delivery of enhanced green fluorescent protein(eGFP) and low cytoto-xicity. This finding is further validated by the cytosolic delivery of RNase A and cytochrome C(Cyt C) to HeLa cells in the viability assay. Together, this work demonstrates that high-throughput screening is an effective and viable approach to the selection of cationic helical polypeptides for the cytosolic delivery of functional proteins.

关键词: Cationic helical polypeptide, Protein delivery, Cytosolic protein

Abstract: Protein delivery is of central importance for both diagnostic and therapeutic applications. However, protein delivery faces challenges including poor endosomal escape and thus limited efficiency. Here, we report the facile construction and screening of a small library of cationic helical polypeptides for cytosolic protein delivery. The library is based on a random copolymer poly(γ-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}esteryl-L-glutamate)-randompoly(γ-6-chlorohexyl-L-glutamate)[P(EG₃-r-ClC₆)Glu], which is then modified with various pyridine derivatives and alkyl thiols. Flow Cytometry, confocal laser scanning microscopy, and viability assay collaboratively identify two leading polymers, showing efficient delivery of enhanced green fluorescent protein(eGFP) and low cytoto-xicity. This finding is further validated by the cytosolic delivery of RNase A and cytochrome C(Cyt C) to HeLa cells in the viability assay. Together, this work demonstrates that high-throughput screening is an effective and viable approach to the selection of cationic helical polypeptides for the cytosolic delivery of functional proteins.

Key words: Cationic helical polypeptide, Protein delivery, Cytosolic protein

WANG Ruijue, SHENG Kai, HOU Yingqin, SUN Jialing, LU Hua. Tailoring Cationic Helical Polypeptides for Efficient Cytosolic Protein Delivery[J]. 高等学校化学研究, 2020, 36(1): 134-138.

WANG Ruijue, SHENG Kai, HOU Yingqin, SUN Jialing, LU Hua. Tailoring Cationic Helical Polypeptides for Efficient Cytosolic Protein Delivery[J]. Chemical Research in Chinese Universities, 2020, 36(1): 134-138.

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Tailoring Cationic Helical Polypeptides for Efficient Cytosolic Protein Delivery

Tailoring Cationic Helical Polypeptides for Efficient Cytosolic Protein Delivery

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