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Exploration of Peptide Inhibitors of Human Squalene Synthase through Molecular Modeling and Phage Display Technique

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Abstract

Many studies have demonstrated the role of elevated levels of serum cholesterol in the pathogenesis of atherosclerosis and coronary heart disease. Various drugs targeting the key enzymes involved in the cholesterol biosynthesis pathway have been investigated for the treatment of hypercholesterolemia. Human squalene synthase has been one of the most important targets for therapeutic intervention. In the present study, we used the recombinant human squalene synthase as the lure for screening the peptide inhibitors from phage-displayed random peptide library. The tightly bound phages and their derived peptides were further evaluated based on their potential binding capabilities, molecular modeling characteristics and predicted absorption, distribution, metabolism, excretion, toxicity (ADMET) properties. Several hexa-peptides and tetra-peptides were finally synthesized to assay their inhibitory effects toward the recombinant human squalene synthase. The results demonstrated that the hexa-peptide FTACNW and tetra-peptide VACL can inhibit human squalene synthase effectively (with IC50 values near 100 μM) and may have potential to develop further as future hypocholesterolemia agents.

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Abbreviations

ADMET:

Absorption, distribution, metabolism, excretion, toxicity

ELISA:

Enzyme-linked immune-sorbent assay

FPP:

Farnesyl diphosphate

hSQS:

Human squalene synthase

HMG-CoA:

3-hydroxy-3-methylglutaryl-coenzyme A

hHMGR:

human HMG-CoA reductase

hHMGRI:

human HMG-CoA reductase inhibitor

IPTG:

Isopropyl β-d-1-thiogalactopyranoside

LDL-R:

Low-density lipoprotein receptor

MTT:

3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide

PCR:

Polymerase chain reaction

PSQPP:

Presqualene diphosphate

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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Acknowledgments

This work was partially supported by grants from National Science Council ROC (NSC97 -2311-B259-04-MY3 to D. Shiuan, 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, Hwan-Kang Lin & Yue-Hao Chen

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

    Ding-Kwo Chang

  3. Development Center for Biotechnology, Taipei, Taiwan, 221, Republic of China

    Kao-Jean Huang

  4. Department of Applied Chemistry, National Pingtung University, Pingtung, Taiwan, 900, Republic of China

    Lynn Farh

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  1. David Shiuan
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  2. Hwan-Kang Lin
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  4. Ding-Kwo Chang
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  5. Kao-Jean Huang
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Correspondence to David Shiuan.

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Shiuan, D., Lin, HK., Chen, YH. et al. Exploration of Peptide Inhibitors of Human Squalene Synthase through Molecular Modeling and Phage Display Technique. Appl Biochem Biotechnol 178, 312–323 (2016). https://doi.org/10.1007/s12010-015-1873-y

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