Limits...
Effects of protein transduction domain (PTD) selection and position for improved intracellular delivery of PTD-Hsp27 fusion protein formulations

View Article: PubMed Central - PubMed

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.

No MeSH data available.


Schematic illustration of transduction of PTD-Hsp27 fusion proteins and their therapeutic effects on apoptosis
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC5037156&req=5

Fig1: Schematic illustration of transduction of PTD-Hsp27 fusion proteins and their therapeutic effects on apoptosis

Mentions: In this study, we intend to prove significance of PTD selection by using PTD-conjugated heat shock protein 27 (Hsp27) fusion proteins. Hsp27 is a chaperone with a molecular weight of 27 kDa. It inhibits apoptosis against hypoxia in cells by interacting directly with caspase activation components (Tan et al. 2009), especially when linked with PTDs, it showed significant effects against hypoxia as shown in Fig. 1 (Kwon et al. 2007; Liu et al. 2014). 9R was conjugated to C-terminus and N-terminus of Hsp27-protein to compare effect of PTDs by their locations. Tat conjugated to Hsp27 was used as a positive control (Tan et al. 2009). As 9R-Hsp27 showed better results than Hsp27-9R, Tat was conjugated at N-terminal of Hsp27-9R to observe improvement in Hsp27-9R. After anti-apoptotic efficacy comparison, it was concluded that N-terminal conjugated PTD-Hsp27 fusion proteins have more protective effects than C-terminal.Fig. 1


Effects of protein transduction domain (PTD) selection and position for improved intracellular delivery of PTD-Hsp27 fusion protein formulations
Schematic illustration of transduction of PTD-Hsp27 fusion proteins and their therapeutic effects on apoptosis
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037156&req=5

Fig1: Schematic illustration of transduction of PTD-Hsp27 fusion proteins and their therapeutic effects on apoptosis
Mentions: In this study, we intend to prove significance of PTD selection by using PTD-conjugated heat shock protein 27 (Hsp27) fusion proteins. Hsp27 is a chaperone with a molecular weight of 27 kDa. It inhibits apoptosis against hypoxia in cells by interacting directly with caspase activation components (Tan et al. 2009), especially when linked with PTDs, it showed significant effects against hypoxia as shown in Fig. 1 (Kwon et al. 2007; Liu et al. 2014). 9R was conjugated to C-terminus and N-terminus of Hsp27-protein to compare effect of PTDs by their locations. Tat conjugated to Hsp27 was used as a positive control (Tan et al. 2009). As 9R-Hsp27 showed better results than Hsp27-9R, Tat was conjugated at N-terminal of Hsp27-9R to observe improvement in Hsp27-9R. After anti-apoptotic efficacy comparison, it was concluded that N-terminal conjugated PTD-Hsp27 fusion proteins have more protective effects than C-terminal.Fig. 1

View Article: PubMed Central - PubMed

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.

No MeSH data available.