Characterization and molecular docking study of cathepsin L inhibitory peptides (SnuCalCpIs) from Calotropis procera R. Br

• Published in: Scientific reports 2022-04, Vol.12 (1), p.5825-5825, Article 5825
• Main Authors: Kwon, Chang Woo, Yeo, Subin, Chang, Pahn-Shick
• Format: Article
• Language: English
• Subjects: 631/45/607/468; 631/45/611; 631/45/612; 631/67/2195; Agricultural biotechnology; Calotropis - metabolism; Calotropis procera; Cancer; Cathepsin L; Cathepsin L - metabolism; Cell migration; E coli; Enzyme Precursors - metabolism; Enzymes; Humanities and Social Sciences; Medicinal plants; Molecular Docking Simulation; multidisciplinary; Peptides; Peptides - metabolism; Plasmids; Proenzymes; Proteins; Science; Science (multidisciplinary); Transcriptomes
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-022-09854-x

Propeptides, released from the autocatalytic activation of its zymogen, are potential inhibitors against proteases involved in cancer cell invasion and migration. Our research team previously obtained novel propeptides (SnuCalCpIs) from transcriptome analysis of the medicinal plant Calotropis procera R. Br. and reported them as promising candidates for cancer therapeutics due to their cathepsin L inhibition activity. In the present study, inhibitory activity among SnuCalCpIs was compared with inhibition efficiency and verified by in silico molecular docking analysis. Only SnuCalCpI03 and SnuCalCpI15, expressed in Escherichia coli , showed inhibitory activity against cathepsin L as competitive inhibitors, and the half-maximal inhibitory concentrations (IC 50 ) values of 2.1 nM and 1.6 nM, respectively. They were stable below 70 °C, maintaining more than 90% inhibitory activity over a wide range of pH (2.0–10.0), except at the isoelectric point (pI). The template-based docking simulation models showed that SnuCalCpI02, SnuCalCpI12, and SnuCalCpI16 could not interact with the substrate-binding cleft of cathepsin L even though they possessed the same conserved domain. In contrast, SnuCalCpI03 and SnuCalCpI15 interacted with cathepsin L along the propeptide binding loop and substrate-binding cleft, resulting in obstruction of substrate access to the active site.