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Isolation and Characterization of a Kunitz-Type Trypsin Inhibitor with Antiproliferative Activity from Gymnocladus Chinensis (Yunnan Bean) Seeds

A 20-kDa Kunitz-type trypsin inhibitor was isolated from Gymnocladus chinensis (Yunnan bean) seeds. The isolation procedure involved ion exchange chromatography on diethylaminoethyl cellulose (DEAE-cellulose), affinity chromatography on Affi-gel blue gel, ion exchange chromatography on sulfopropyl s...

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Detalles Bibliográficos
Autores principales: Zhu, M. J., Zhang, G. Q., Wang, H. X., Ng, T. B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer US 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7088384/
https://www.ncbi.nlm.nih.gov/pubmed/21468674
http://dx.doi.org/10.1007/s10930-011-9325-1
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author Zhu, M. J.
Zhang, G. Q.
Wang, H. X.
Ng, T. B.
author_facet Zhu, M. J.
Zhang, G. Q.
Wang, H. X.
Ng, T. B.
author_sort Zhu, M. J.
collection PubMed
description A 20-kDa Kunitz-type trypsin inhibitor was isolated from Gymnocladus chinensis (Yunnan bean) seeds. The isolation procedure involved ion exchange chromatography on diethylaminoethyl cellulose (DEAE-cellulose), affinity chromatography on Affi-gel blue gel, ion exchange chromatography on sulfopropyl sepharose (SP-sepharose), and gel filtration by FPLC on Superdex 75. The trypsin inhibitor was adsorbed on DEAE-cellulose, unadsorbed on Affi-gel blue gel, and adsorbed on SP-Sepharose. It dose-dependently inhibited trypsin with an IC(50) value of 0.4 μM. Dithiothreitol reduced its trypsin inhibitory activity, suggesting that an intact disulfide bond is indispensable to the activity. It suppressed [methyl-(3)H] thymidine incorporation by leukemia L1210 cells and lymphoma MBL2 cells with an IC(50) value of 4.7 and 9.4 μM, respectively. There was no effect on human immunodeficiency virus(4)-1 reverse transcriptase activity and fungal growth when the trypsin inhibitor was tested up to 100 μM.
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spelling pubmed-70883842020-03-23 Isolation and Characterization of a Kunitz-Type Trypsin Inhibitor with Antiproliferative Activity from Gymnocladus Chinensis (Yunnan Bean) Seeds Zhu, M. J. Zhang, G. Q. Wang, H. X. Ng, T. B. Protein J Article A 20-kDa Kunitz-type trypsin inhibitor was isolated from Gymnocladus chinensis (Yunnan bean) seeds. The isolation procedure involved ion exchange chromatography on diethylaminoethyl cellulose (DEAE-cellulose), affinity chromatography on Affi-gel blue gel, ion exchange chromatography on sulfopropyl sepharose (SP-sepharose), and gel filtration by FPLC on Superdex 75. The trypsin inhibitor was adsorbed on DEAE-cellulose, unadsorbed on Affi-gel blue gel, and adsorbed on SP-Sepharose. It dose-dependently inhibited trypsin with an IC(50) value of 0.4 μM. Dithiothreitol reduced its trypsin inhibitory activity, suggesting that an intact disulfide bond is indispensable to the activity. It suppressed [methyl-(3)H] thymidine incorporation by leukemia L1210 cells and lymphoma MBL2 cells with an IC(50) value of 4.7 and 9.4 μM, respectively. There was no effect on human immunodeficiency virus(4)-1 reverse transcriptase activity and fungal growth when the trypsin inhibitor was tested up to 100 μM. Springer US 2011-04-06 2011 /pmc/articles/PMC7088384/ /pubmed/21468674 http://dx.doi.org/10.1007/s10930-011-9325-1 Text en © Springer Science+Business Media, LLC 2011 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Article
Zhu, M. J.
Zhang, G. Q.
Wang, H. X.
Ng, T. B.
Isolation and Characterization of a Kunitz-Type Trypsin Inhibitor with Antiproliferative Activity from Gymnocladus Chinensis (Yunnan Bean) Seeds
title Isolation and Characterization of a Kunitz-Type Trypsin Inhibitor with Antiproliferative Activity from Gymnocladus Chinensis (Yunnan Bean) Seeds
title_full Isolation and Characterization of a Kunitz-Type Trypsin Inhibitor with Antiproliferative Activity from Gymnocladus Chinensis (Yunnan Bean) Seeds
title_fullStr Isolation and Characterization of a Kunitz-Type Trypsin Inhibitor with Antiproliferative Activity from Gymnocladus Chinensis (Yunnan Bean) Seeds
title_full_unstemmed Isolation and Characterization of a Kunitz-Type Trypsin Inhibitor with Antiproliferative Activity from Gymnocladus Chinensis (Yunnan Bean) Seeds
title_short Isolation and Characterization of a Kunitz-Type Trypsin Inhibitor with Antiproliferative Activity from Gymnocladus Chinensis (Yunnan Bean) Seeds
title_sort isolation and characterization of a kunitz-type trypsin inhibitor with antiproliferative activity from gymnocladus chinensis (yunnan bean) seeds
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7088384/
https://www.ncbi.nlm.nih.gov/pubmed/21468674
http://dx.doi.org/10.1007/s10930-011-9325-1
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