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Discovering Peptide Inhibitors of Human Squalene Synthase Through Screening the Phage-Displayed Cyclic Peptide c7c Library

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Abstract

Many drugs for the treatment of hypercholesterolemia are targeting the enzymes involved in human cholesterol biosynthesis pathway. Squalene synthase, the rate-limiting enzyme located at the downstream of cholesterol synthesis pathway, has become a better candidate to develop next-generation hypocholesterolemia drugs. In the present study, we cloned and expressed the recombinant human squalene synthase (hSQS) as the lure to isolate potential peptide inhibitors from screening the conformation-constrained phage-displayed cyclic peptide c7c library. Their binding capabilities were further estimated by ELISA. Their pharmaceutical potentials were then analyzed through molecular modeling and the ADMET property evaluations. Four ennea-peptides and nine tetra-peptides were finally synthesized to evaluate their inhibitory potentials toward hSQS. The results indicate that the ennea-peptide CLSPHSMFC, tetra-peptides SMFC, CKTE, and WHQW can effectively inhibit hSQS activities (IC50 values equal to 64, 76, 87, and 90 μM, respectively). These peptides may have potentials to develop future cholesterol-lowering therapeutics. The ligand-protein interaction analysis also reveals that the inner hydrophobic pocket could be a more critical site of hSQS.

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Abbreviations

ELISA:

Enzyme-linked immunosorbent assay

hHMGR:

Human 3-hydroxy-3-methylglutaryl-coenzyme A reductase

hSQS:

Human squalene synthase

IPTG:

Isopropyl β-d-1-thiogalactopyranoside

LDL-R:

Low-density lipoprotein receptor

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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Acknowledgments

This work was partially supported by grants from the Ministry of Science and Technology, Taiwan, ROC (NSC97-2311-B259-04-MY3 to D. Shiuan, MOST 103-2113-M-259-019 to D. F. Tai and NSC99-2113-M-001-024-MY3 to D. K. Chang.).

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Authors and Affiliations

  1. Department of Life Science and Institute of Biotechnology, National Dong Hwa University, Hualien, Taiwan, 974, Republic of China

    David Shiuan, Yue-Hao Chen, Hwan-Kang Lin & Kao-Jean Huang

  2. Development Center for Biotechnology, Taipei, Taiwan, 115, Republic of China

    Kao-Jean Huang

  3. Department of Chemistry, National Dong Hwa University, Hualien, Taiwan, 974, Republic of China

    Da-Fu Tai

  4. Institute of Chemistry, Academia Sinica, Taipei, Taiwan, 115, Republic of China

    Ding-Kwo Chang

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  1. David Shiuan
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  2. Yue-Hao Chen
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  3. Hwan-Kang Lin
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  4. Kao-Jean Huang
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  5. Da-Fu Tai
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  6. Ding-Kwo Chang
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Correspondence to David Shiuan.

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Shiuan, D., Chen, YH., Lin, HK. et al. Discovering Peptide Inhibitors of Human Squalene Synthase Through Screening the Phage-Displayed Cyclic Peptide c7c Library. Appl Biochem Biotechnol 179, 597–609 (2016). https://doi.org/10.1007/s12010-016-2016-9

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  • DOI: https://doi.org/10.1007/s12010-016-2016-9

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