title: Discovery of Anti-Hypertensive Oligopeptides from Adlay Based on In Silico Proteolysis and Virtual Screening.
abstract: Adlay (Coix larchryma-jobiL.) was the commonly used Traditional Chinese Medicine (TCM) with high content of seed storage protein. The hydrolyzed bioactive oligopeptides of adlay have been proven to be anti-hypertensive effective components. However, the structures and anti-hypertensive mechanism of bioactive oligopeptides from adlay were not clear. To discover the definite anti-hypertensive oligopeptides from adlay, in silico proteolysis and virtual screening were implemented to obtain potential oligopeptides, which were further identified by biochemistry assay and molecular dynamics simulation. In this paper, ten sequences of adlay prolamins were collected and in silico hydrolyzed to construct the oligopeptide library with 134 oligopeptides. This library was reverse screened by anti-hypertensive pharmacophore database, which was constructed by our research team and contained ten anti-hypertensive targets. Angiotensin-I converting enzyme (ACE) was identified as the main potential target for the anti-hypertensive activity of adlay oligopeptides. Three crystal structures of ACE were utilized for docking studies and 19 oligopeptides were finally identified with potential ACE inhibitory activity. According to mapping features and evaluation indexes of pharmacophore and docking, three oligopeptides were selected for biochemistry assay. An oligopeptide sequence, NPATY (IC50= 61.88 ± 2.77 µM), was identified as the ACE inhibitor by reverse-phase high performance liquid chromatography (RP-HPLC) assay. Molecular dynamics simulation of NPATY was further utilized to analyze interactive bonds and key residues. ALA354 was identified as a key residue of ACE inhibitors. Hydrophobic effect of VAL518 and electrostatic effects of HIS383, HIS387, HIS513 and Zn2+were also regarded as playing a key role in inhibiting ACE activities. This study provides a research strategy to explore the pharmacological mechanism of Traditional Chinese Medicine (TCM) proteins based on in silico proteolysis and virtual screening, which could be beneficial to reveal the pharmacological action of TCM proteins and provide new lead compounds for peptides-based drug design.
authors.affiliation: Key Laboratory of TCM Foundation and New Drug Research, School of Chinese Material Medica, University of Chinese Medicine, Beijing 100102, China. b20100222012@163.com.
authors.forename: Liansheng
authors.initials: L
authors.lastname: Qiao
authors.affiliation: Key Laboratory of TCM Foundation and New Drug Research, School of Chinese Material Medica, University of Chinese Medicine, Beijing 100102, China. libinzyy@163.com.
authors.forename: Bin
authors.initials: B
authors.lastname: Li
authors.affiliation: Key Laboratory of TCM Foundation and New Drug Research, School of Chinese Material Medica, University of Chinese Medicine, Beijing 100102, China. 18811791975@163.com.
authors.forename: Yankun
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Key Laboratory of TCM Foundation and New Drug Research, School of Chinese Material Medica, University of Chinese Medicine, Beijing 100102, China. lilingling94@126.com.
authors.forename: Lingling
authors.initials: L
authors.lastname: Li
authors.affiliation: Key Laboratory of TCM Foundation and New Drug Research, School of Chinese Material Medica, University of Chinese Medicine, Beijing 100102, China. chenxi_cx95@163.com.
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
authors.affiliation: Key Laboratory of TCM Foundation and New Drug Research, School of Chinese Material Medica, University of Chinese Medicine, Beijing 100102, China. wanglz@bucm.edu.cn.
authors.forename: Lingzhi
authors.initials: L
authors.lastname: Wang
authors.affiliation: Key Laboratory of TCM Foundation and New Drug Research, School of Chinese Material Medica, University of Chinese Medicine, Beijing 100102, China. lufang1017@163.com.
authors.forename: Fang
authors.initials: F
authors.lastname: Lu
authors.affiliation: Key Laboratory of TCM Foundation and New Drug Research, School of Chinese Material Medica, University of Chinese Medicine, Beijing 100102, China. 17801080765@163.com.
authors.forename: Ganggang
authors.initials: G
authors.lastname: Luo
authors.affiliation: Key Laboratory of TCM Foundation and New Drug Research, School of Chinese Material Medica, University of Chinese Medicine, Beijing 100102, China. lidoc2727@163.com.
authors.forename: Gongyu
authors.initials: G
authors.lastname: Li
authors.affiliation: Key Laboratory of TCM Foundation and New Drug Research, School of Chinese Material Medica, University of Chinese Medicine, Beijing 100102, China. zhangyanling@bucm.edu.cn.
authors.forename: Yanling
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000806:Angiotensin-Converting Enzyme Inhibitors / Q000737:chemistry; D000959:Antihypertensive Agents / Q000494:pharmacology*; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D031742:Coix / Q000737:chemistry*; D003198:Computer Simulation; D018360:Crystallography, X-Ray; D030562:Databases, Protein; D055808:Drug Discovery; D004353:Drug Evaluation, Preclinical; D062105:Molecular Docking Simulation; D009842:Oligopeptides / Q000494:pharmacology*; D019151:Peptide Library; D059748:Proteolysis; D013997:Time Factors; D014584:User-Computer Interface
chemical_list: D000806:Angiotensin-Converting Enzyme Inhibitors; D000959:Antihypertensive Agents; D009842:Oligopeptides; D019151:Peptide Library
keywords: adlay; angiotensin-I converting enzyme (ACE); hypertension; in silico proteolysis; molecular dynamics (MD); oligopeptides