title: Effects of protein transduction domain (PTD) selection and position for improved intracellular delivery of PTD-Hsp27 fusion protein formulations.
abstract: Protein drugs have attracted considerable attention as therapeutic agents due to their diversity and biocompatibility. However, hydrophilic proteins possess difficulty in penetrating lipophilic cell membrane. Although protein transduction domains (PTDs) have shown effectiveness in protein delivery, the importance of selection and position of PTDs in recombinant protein vector constructs has not been investigated. This study intends to investigate the significance of PTD selection and position for therapeutic protein delivery. Heat shock protein 27 (Hsp27) would be a therapeutic protein for the treatment of ischemic heart diseases, but itself is insufficient to prevent systemic degradation and overcoming biochemical barriers during cellular transport. Among all PTD-Hsp27 fusion proteins we cloned, Tat-Hsp27 fusion protein showed the highest efficacy. Nona-arginine (9R) conjugation to the N-terminal of Hsp27 (Hsp27-T) showed higher efficacy than C-terminal. To test the synergistic effect of two PTDs, Tat was inserted to the N-terminal of Hsp27-9R. Tat-Hsp27-9R exhibited enhanced transduction efficiency and significant improvement against oxidative stress and apoptosis. PTD-Hsp27 fusion proteins have strong potential to be developed as therapeutic proteins for the treatment of ischemic heart diseases and selection and position of PTDs for improved efficacy of PTD-fusion proteins need to be optimized considering protein's nature, transduction efficiency and stability.
authors.affiliation: Department of Bioengineering, Institute for Bioengineering and Biopharmaceutical Research, Hanyang University, 17 Haengdang-dong, Seongdong-gu, Seoul, 04763, Korea.
authors.forename: Qurrat
authors.initials: Q
authors.lastname: Ul Ain
authors.affiliation: Department of Bioengineering, Institute for Bioengineering and Biopharmaceutical Research, Hanyang University, 17 Haengdang-dong, Seongdong-gu, Seoul, 04763, Korea.
authors.forename: Jong Hwan
authors.initials: JH
authors.lastname: Lee
authors.affiliation: Department of Bioengineering, Institute for Bioengineering and Biopharmaceutical Research, Hanyang University, 17 Haengdang-dong, Seongdong-gu, Seoul, 04763, Korea.
authors.forename: Young Sun
authors.initials: YS
authors.lastname: Woo
authors.affiliation: Department of Bioengineering, Institute for Bioengineering and Biopharmaceutical Research, Hanyang University, 17 Haengdang-dong, Seongdong-gu, Seoul, 04763, Korea. yongheekim@hanyang.ac.kr.|BK 21 Plus Future Biopharmaceutical Human Resources Training and Research Team, Department of Bioengineering, Institute for Bioengineering and Biopharmaceutical Research, Hanyang University, 17 Haengdang-dong, Seongdong-gu, Seoul, 04763, Korea. yongheekim@hanyang.ac.kr.
authors.forename: Yong-Hee
authors.initials: YH
authors.lastname: Kim
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004339:Drug Compounding / Q000379:methods; D016503:Drug Delivery Systems / Q000379:methods*; D055551:HSP27 Heat-Shock Proteins / Q000008:administration & dosage* / Q000378:metabolism; D006801:Humans; D007424:Intracellular Fluid / Q000187:drug effects* / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism; D000072417:Protein Domains; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage* / Q000378:metabolism
chemical_list: D055551:HSP27 Heat-Shock Proteins; D011993:Recombinant Fusion Proteins
keywords: Fusion proteins; Heat shock protein 27; Hypoxia; Selection and position of protein transduction domains (PTDs)