Revealing the Sequence Characteristics and Molecular Mechanisms of ACE Inhibitory Peptides by Comprehensive Characterization of 160,000 Tetrapeptides

Abstract

Chronic diseases, such as hypertension, cause great harm to human health. Conventional drugs have promising therapeutic effects, but also cause significant side effects. Food-sourced angiotensin-converting enzyme (ACE) inhibitory peptides are an excellent therapeutic alternative to pharmaceuticals, as they have fewer side effects. However, there is no systematic and effective screening method for ACE inhibitory peptides, and the lack of understanding of the sequence characteristics and molecular mechanism of these inhibitory peptides poses a major obstacle to the development of ACE inhibitory peptides. Through systematically calculating the binding effects of 160,000 tetrapeptides with ACE by molecular docking, we found that peptides with Tyr, Phe, His, Arg, and especially Trp were the characteristic amino acids of ACE inhibitory peptides. The tetrapeptides of WWNW, WRQF, WFRV, YYWK, WWDW, and WWTY rank in the top 10 peptides exhibiting significantly high ACE inhibiting behaviors, with IC50 values between 19.98 ± 8.19 μM and 36.76 ± 1.32 μM. Salt bridges, π–π stacking, π–cations, and hydrogen bonds contributed to the high binding characteristics of the inhibitors and ACE. Introducing eight Trp into rabbit skeletal muscle protein (no Trp in wide sequence) endowed the protein with a more than 90% ACE inhibition rate, further suggesting that meat with a high content of Trp could have potential utility in hypertension regulation. This study provides a clear direction for the development and screening of ACE inhibitory peptides.

Two Novel Angiotensin-Converting Enzyme (ACE) Inhibitory and ACE2 Upregulating Peptides from the Hydrolysate of Pumpkin (Cucurbita moschata) Seed Meal

“…The three major active site pockets of ACE are S1, S2, and S1′ . ACE inhibitory peptides are connected to ACE through interactions such as van der Waals forces, hydrogen bonding (H-bonds), hydrophobic forces, and electrostatic forces, among which hydrogen bonding is the major force that maintains the binding of ACE to the inhibitory peptides . In addition, Zn 2+ is also an active site of ACE, forming a tetrahedral coordination structure with ACE residues His383, His387, and Glu411, which partially contributes to the binding strength between ACE and its inhibitors .…”

Preparation, identification, and molecular docking of novel angiotensin‐converting enzyme inhibitory peptides derived from rice‐based distillers' spent cakes

“…Therefore, we speculated that docking with the ACE active site may be responsible for the higher ACE inhibitory activity of the short peptides, whereas the binding of medium‐sized peptides to the inactive binding sites of ACE, such as Glu123, Asp121, Arg522, and Lys118, could have increased the ACE inhibitory ability of the peptides. Simultaneously, salt bridges, pi‐pi stacking and pi‐cations favor the tight binding of ACE inhibitors 48 …”

Targeted Affinity Purification and Mechanism of Action of Angiotensin-Converting Enzyme (ACE) Inhibitory Peptides from Sea Cucumber Gonads

“…The peptides' Grand average of hydropathicity (GRAVY) and toxicity were computed using ToxinPred (https://webs.iiitd.edu.in/ raghava/toxinpred/ (accessed on 8 June 2023)) [43]. The pharmacokinetic properties of the peptides were analyzed using the online tool ADMETlab (https://admetmesh.scbdd.com/ (accessed on 9 June 2023)), focusing on indicators such as human intestinal absorption (HIA), blood-brain barrier (BBB) penetration, CYP450 metabolism inhibition parameters, and acute oral toxicity [14]. The ACE inhibitory activities of the 20 peptides were determined at a concentration of 1 mg/mL, and the peptides with superior activities were selected.…”

“…We also performed a molecular dynamics (MD) simulation and binding free energy analysis to validate these interactions, providing a deeper understanding of the binding process and conformational changes within the complex system. When these methods are combined with the prediction of ADMET properties, they offer a fast and efficient means of screening potential drugs [14]. Finally, the feasibility of this approach was verified by analyzing the in vivo antihypertensive activity in SHRs.…”