Submit Manuscript  

Article Details


Expression and Purification of Delta Sleep-Inducing Peptide Fused with Protein Transduction Domain and Human Serum Albumin in Pichia pastoris

[ Vol. 24 , Issue. 7 ]

Author(s):

Xin-Guo Zhang*, Wen-Na Wang, Chun-Sheng Zhang, Kun Li, Guo-Di Ma and Jian-Yong Li   Pages 668 - 675 ( 8 )

Abstract:


Background: Sleep is a natural part of every individual’s life. Delta sleep-inducing peptide (DSIP) is a nonapeptide that could promote sleep through the induction of slow wave sleep. However, little is known about the pharmacological effect of DSIP on insomnia.

Objectives: The main objective of this study was to analyze the pharmacological effect of DSIP on insomnia.

Methods: We designed a fusion protein containing N-terminal TAT-based transduction domain followed by human serum albumin and DSIP and designated this protein as PHD fusion protein. The PHD fusion protein were expressed in Pichia pastoris and purified. Mice were administered single subcutaneous injections three concentrations of PHD fusion protein (0.5, 1, 2 mg/kg), and the pharmacological activity of PHD fusion protein was studied using classic pentobarbitalinduced sleep test.

Results: We expressed the PHD fusion protein in P. pastoris; furthermore, the PHD fused protein was purified to near homogeneity by DEAE Sepharose FF, Phenyl Sepharose HP and Blue Sepharose 6 FF. Our result showed that the increase of pentobarbital-induced hypnotic effect characterized by reducing sleep latency and prolonged sleep duration was observed for increasing concentrations of PHD fusion protein (P<0.05); moreover, different dose of PHD fusion protein could induce the mice to re-sleep in a dose-dependent manner, whereas higher doses of PHD fusion protein (1.0, 2.0 mg/kg) significantly increased the rate of sleep re-onset compared with the vehicle group of mice (P<0.05).

Conclusion: PHD fusion protein increased the hypnotic effects of pentobarbital by reducing sleep latency and prolonged sleep duration. The present study suggested PHD fusion protein could be a new drug candidate for insomnia.

Keywords:

Delta sleep-inducing peptide, human serum albumin, protein transduction domain, Pichia pastoris, fusion protein.

Affiliation:

School of Life Science and Engineering, Lanzhou University of Technology, No.287 Langongping Road, Qilihe District, Lanzhou City, Gansu Province, School of Life Science and Engineering, Lanzhou University of Technology; Key Laboratory of Screening and Processing in new Tibetan medicine of Gansu Province, Gansu, 730050, School of Life Science and Engineering, Lanzhou University of Technology; Key Laboratory of Screening and Processing in new Tibetan medicine of Gansu Province, Gansu, 730050, School of Life Science and Engineering, Lanzhou University of Technology; Key Laboratory of Screening and Processing in new Tibetan medicine of Gansu Province, Gansu, 730050, School of Life Science and Engineering, Lanzhou University of Technology; Key Laboratory of Screening and Processing in new Tibetan medicine of Gansu Province, Gansu, 730050, Key Lab of New Animal Drug Project, Gansu Province; Key Lab of Veterinary Pharmaceutical Development, Ministry of Agriculture; Lanzhou Institute of Husbandry and Pharmaceutical Sciences of CAAS, Lanzhou, Gansu 730050

Graphical Abstract:



Read Full-Text article