Identification and pharmaceutical evaluation of novel frog skin-derived serine proteinase inhibitor peptide–PE-BBI (Pelophylax esculentus Bowman-Birk inhibitor) for the potential treatment of cancer

Abstract

Amphibian venom-derived peptides have high potential in the field of anticancer drug discovery. We have isolated a novel Bowman-Birk proteinase inhibitor (BBI)-type peptide from the skin secretion of Pelophylax esculentus (PE) named PE-BBI, and evaluated its bio-functions and anti-cancer activity in vitro. PE-BBI is a heptadecapeptide with C-terminal amidation. The mRNA sequence and primary structure of PE-BBI were identified using RT-PCR and LC/MS, respectively. A trypsin inhibitory assay was used to characterize the serine proteinase inhibitory activity of synthetic PE-BBI. PE-BBI’s myotropic activity was analyzed using isolated rat bladder and rat-tail artery smooth muscle tissues, and the anti-cancer ability of PE-BBI using human colorectal cancer cells. PE-BBI’s mechanism of action was investigated using Discovery studio software. PE-BBI showed trypsin inhibitory activity (Ki = 310 ± 72 nM), strong myotropic activity, and cytotoxicity that were specific to cancer cells, and no side effect to normal epithelial cells. The docking stimulation showed that PE-BBI had high affinity to several members of human kallikrein related peptidase (KLK) family. This finding helps to enrich our understanding of BBI peptides’ mode of action. Moreover, the data presented here validates frog secretions as sources of potential novel proteinase inhibitors for cancer treatment.

Discovery and Rational Design of a Novel Bowman-Birk Related Protease Inhibitor

“…In the report by Zhang et al [12], a Bowman-Birk type chymotrypsin inhibitory peptide, HECI, and its analogue, K 9 -HECI, exhibited great anti-proliferative potency against H157, PC-3 and MCF-7. Also, PE-BBI was reported to have an inhibition effect on several colon cancer cell lines [13]. In our study, Tat-loop suppresses the growth of the lung cancer cells H460 and H157.…”

“…The skin of frogs is the main organ involved in their defense system, which manufactures diverse bioactive peptides that possess cytolytic pharmacological activities [6], and as a result it is also an excellent source of protease inhibitors. To date, many BBIs isolated from amphibians have been reported, such as peptide leucine arginine (pLR) [7], peptide tyrosine arginine (pYR) [8], the Bowman-Birk-like trypsin inhibitor from Huia versabilis (HV-BBI) [9], Hejiang trypsin inhibitor (HJTI) [10], the Bowman-Birk-type inhibitor from Odorrana schmackeri (OSTI) [11], Hylarana erythraea chymotrypsin inhibitor (HECI) [12] and Pelophyla esculentus Bowman-Birk proteinase inhibitor (PE-BBI) [13]. Generally, the BBI peptides from amphibians possess a highly-conserved 11-residue canonical disulfide loop, which is different from plant BBIs.…”

Clonagem Molecular Da L-Asparaginase De Proteus Vulgaris Em Vetor De Expressão Para Fusão À Proteína Sumo

“…In the search for alternatives for the treatment of pathologies that have serine proteases, elastase, proteinase 3 and cathepsin G in their pathological processes, a field of studies focused on natural products has shown promising. Some studies report the presence of elastase inhibitors, for example, in seeds of plants of different classes (LYU et al, 2018;RIBEIRO et al, 2010;ROCCO et al, 2011;SUMIKAWA et al, 2006). These inhibitors can be used in the clinic in many ways to prevent or treat diseases related to disorders in the inflammatory process.…”

LFB: A Novel Antimicrobial Brevinin-Like Peptide from the Skin Secretion of the Fujian Large Headed Frog, Limnonectes fujianensi

“…According to the study, it was found that numerous AMPs are present among many species of amphibians, presumably for defense against pathogenic organisms [10,11]. The broad-spectrum antimicrobial activities can be chemically synthesized readily; these peptides have become the focus of significant research, spanning some 30 years now [12,13,14,15]. Due to their diversities of primary and secondary structural characters, AMPs from this source have been classified into several specific, structurally-related peptide families.…”