A Novel Trypsin Kunitz-Type Inhibitor from Cajanus cajan Leaves and Its Inhibitory Activity on New Cancer Serine Proteases and Its Effect on Tumor Cell Growth.

Autor: Teixeira EMGF; Departament of Natural Products, Institute of Pharmaceuticals Technology, FIOCRUZ, Av. Brasil 4365, Rio de Janeiro, Rio de Janeiro, 21045-900, Brazil.; Laboratory of Bioprospection and Applied Ethnopharmacology, Federal University of Rio de Janeiro, Rio de Janeiro, 21941-902, Brazil., Kalume DE; Interdisciplinary Laboratory of Medical Research, IOC-Oswaldo Cruz Institute, FIOCRUZ, Av. Brasil 4365, Rio de Janeiro, Rio de Janeiro, CEP 21045-900, Brazil., Ferreira PF; Departament of Natural Products, Institute of Pharmaceuticals Technology, FIOCRUZ, Av. Brasil 4365, Rio de Janeiro, Rio de Janeiro, 21045-900, Brazil., Alves TA; Departament of Natural Products, Institute of Pharmaceuticals Technology, FIOCRUZ, Av. Brasil 4365, Rio de Janeiro, Rio de Janeiro, 21045-900, Brazil., Fontão APGA; Departament of Pharmacology, Institute of Pharmaceuticals Technology, FIOCRUZ, Av. Brasil 4365, Rio de Janeiro, Rio de Janeiro, CEP 21045-900, Brazil., Sampaio ALF; Departament of Pharmacology, Institute of Pharmaceuticals Technology, FIOCRUZ, Av. Brasil 4365, Rio de Janeiro, Rio de Janeiro, CEP 21045-900, Brazil., de Oliveira DR; Laboratory of Bioprospection and Applied Ethnopharmacology, Federal University of Rio de Janeiro, Rio de Janeiro, 21941-902, Brazil., Morgado-Díaz JA; Cellular and Molecular Oncobiology Program, National Institute of Cancer (INCa), Rio de Janeiro, Brazil., Silva-López RE; Departament of Natural Products, Institute of Pharmaceuticals Technology, FIOCRUZ, Av. Brasil 4365, Rio de Janeiro, Rio de Janeiro, 21045-900, Brazil. raquel.lopez@fiocruz.br.
Jazyk: angličtina
Zdroj: The protein journal [Protein J] 2024 Apr; Vol. 43 (2), pp. 333-350. Date of Electronic Publication: 2024 Feb 12.
DOI: 10.1007/s10930-023-10175-9
Abstrakt: A novel trypsin inhibitor from Cajanus cajan (TIC) fresh leaves was partially purified by affinity chromatography. SDS-PAGE revealed one band with about 15 kDa with expressive trypsin inhibitor activity by zymography. TIC showed high affinity for trypsin (Ki = 1.617 μM) and was a competitive inhibitor for this serine protease. TIC activity was maintained after 24 h of treatment at 70 °C, after 1 h treatments with different pH values, and β-mercaptoethanol increasing concentrations, and demonstrated expressive structural stability. However, the activity of TIC was affected in the presence of oxidizing agents. In order to study the effect of TIC on secreted serine proteases, as well as on the cell culture growth curve, SK-MEL-28 metastatic human melanoma cell line and CaCo-2 colon adenocarcinoma was grown in supplemented DMEM, and the extracellular fractions were submitted salting out and affinity chromatography to obtain new secreted serine proteases. TIC inhibited almost completely, 96 to 89%, the activity of these serine proteases and reduced the melanoma and colon adenocarcinoma cells growth of 48 and 77% respectively. Besides, it is the first time that a trypsin inhibitor was isolated and characterized from C. cajan leaves and cancer serine proteases were isolated and partial characterized from SK-MEL-28 and CaCo-2 cancer cell lines. Furthermore, TIC shown to be potent inhibitor of tumor protease affecting cell growth, and can be one potential drug candidate to be employed in chemotherapy of melanoma and colon adenocarcinoma.
(© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze: MEDLINE
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