Huang, Ju-Hsuan, Nhung Thi Phuong Nong, and Jue-Liang Hsu. “An Efficient Peptidomics Screening for Exogenous Substrates and Inhibitory Peptides of the Dipeptidase ACE from Milk Hydrolysate.” Pharmaceutics 15.2 (2023): 425. https://doi.org/10.3390/pharmaceutics15020425

Abstract

The dipeptidase angiotensin-I-converting enzyme (ACE) pre-incubation, liquid chromatography- mass spectrometry (LC-MS), and stable-isotope labeling were integrated for an efficient screening of ACE’s exogenous substrates from milk hydrolysate. Using this approach, 31 substrates were readily identified from 478 identified peptides and their activities were confirmed using synthetic peptides. Their reactivity is highly correlated with the decreased isotope ratio observed in LC-MS. Among these substrates, the most frequently observed residue at the P1′ position was Leu/Ser. It also revealed that ACE would not cleave the peptide when P1′ is Pro, P2′ is Asp/Glu, or P1 position is Ile. Interestingly, the sequential two-stage hydrolysis was also found. Moreover, their protective effects against ACE-mediated hydrolysis of angiotensin I (Ang-I) were also examined. The result indicated that AYFYPELFR and HLPLPLLQSW can significantly retard the hydrolysis of Ang-I and act as substrate-type inhibitors.