Isolation and functional diversity of Bowman–Birk type serine proteinase inhibitors from Hyacinthus orientalis. Issue 6 (26th March 2021)

• Record Type: Journal Article
• Authors: Aoki-Shioi, Narumi; Terada, Shigeyuki; Hellinger, Roland; Furuta, Yoshitaka; Gruber, Christian W.
• Is Part Of: Biochemical journal. Volume 478:Issue 6(2021)
• Page Start: 1287
• Page End: 1301
• Publication Date: 2021-03-26
• Subjects: cysteine-rich -- lily -- plant peptide -- precursor gene -- serine proteinase inhibitor; Biochemistry -- Periodicals; 572
• DOI: 10.1042/BCJ20201005
• Languages: English
• ISSNs: 0264-6021

Abstract

Abstract : Bowman–Birk inhibitors (BBIs) are plant-derived serine proteinase inhibitors. Endogenously, they function as defense molecules against pathogens and insects, but they also have been explored for applications in cancer treatment and inflammatory disorders. Here, we isolated 15 novel BBIs from the bulb of Hyacinthus orientalis (termed HOSPIs). These isoinhibitors consisted of two or three chains, respectively, that are linked by disulfides bonds based on proposed cleavage sites in the canonical BBI reactive site loop. They strongly inhibited trypsin ( Ki = 0.22–167 nM) and α-chymotrypsin ( Ki = 19–1200 nM). Notably, HOSPI-B4 contains a six-residue reactive loop, which appears to be the smallest such motif discovered in BBIs to date. HOSPI-A6 and -A7 contain an unusual reactive site, i.e. Leu–Met at the P1–P1′ position and have strong inhibitory activity against trypsin, α-chymotrypsin, and elastase. Analysis of the cDNA encoding HOSPIs revealed that the precursors have HOSPI-like domains repeated at least twice with a defined linker sequence connecting individual domains. Lastly, mutational analysis of HOSPIs suggested that the linker sequence does not affect the inhibitory activity, and a Thr residue at the P2 site and a Pro at the P3′ site are crucial for elastase inhibition. Using mammalian proteases as representative model system, we gain novel insight into the sequence diversity and proteolytic activity of plant BBI. These results may aid the rational design of BBI peptides with potent and distinct inhibitory activity against human, pathogen, or insect serine proteinases.