Discovery of Anti-Hypertensive Oligopeptides from Adlay Based on In Silico Proteolysis and Virtual Screening

Abstract

Adlay (Coix larchryma-jobi L.) 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.

Novel ACE inhibitors derived from soybean proteins using in silico and in vitro studies

“…Nowadays, with the development of peptidomics, virtual screening approaches have been utilized to identify ACE inhibitory peptides from food stuff, such as fish (Yu, Chen, et al, ), barley (Gangopadhyay et al, ), adlay (Qiao et al, ), and meat proteins (Lafarga, O'Connor, & Hayes, ), which can simplify the processes, cut down costs, and remain stable. Soybean is widely used as processed foods in food industry.…”

“…The classical approaches to obtain the ACE inhibitory peptides involve protein and enzyme selection, proteolysis of protein, in vitro activity assay, and fractionation, while the main drawback of classical method was time-consuming and sensitivity reduction. Nowadays, with the development of peptidomics, virtual screening approaches have been utilized to identify ACE inhibitory peptides from food stuff, such as fish , barley (Gangopadhyay et al, 2016), adlay (Qiao et al, 2016), and meat proteins (Lafarga, O'Connor, & Hayes, 2014), which can simplify the processes, cut down costs, and remain stable. Soybean is widely used as processed foods in food industry.…”

Novel Angiotensin‐Converting Enzyme Inhibitory Peptides Derived from Oncorhynchus mykiss Nebulin: Virtual Screening and In Silico Molecular Docking Study

“…Nowadays, in silico tools were used to identify food‐derived bioactive peptides in emerging nutraceutical field, including smooth‐hound viscera protein (Abdelhedi et al., ), ACE inhibitory peptide from silkworm pupa (Tao et al., ), prolyl endopeptidase inhibitors from meat (Lafarga, O'Connor, & Hayes, ), antioxidative collagen peptides from carp swim bladder (Roy, Nair, Sen, Soni, & Madhusudhan, ), and antihypertensive oligopeptides from adlay (Qiao et al., ). Oncorhynchus mykiss is a marine fish, and it was highly domesticated, fast‐growing, and an attractive food protein source (Hauge et al., ; Leung et al., ).…”

Using Coexpression Protein Interaction Network Analysis to Identify Mechanisms of Danshensu Affecting Patients with Coronary Heart Disease

“…The proteins corresponding to pharmacophore models with Fit value >0.7 were selected as source proteins used in the following study [42]. …”