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

Abstract

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.

A thermostable trypsin inhibitor with antiproliferative activity from small pinto beans

“…Hence, treatment with DTT would inhibit both types of trypsin inhibitor. On the other hand, most legume Kunitz-type trypsin inhibitors are 20-kDa [34][35][36][37][38] while legume BBIs are 16-kDa in molecular weight 27,39 . In this report, the small pinto bean trypsin inhibitor was found to be a 16-kDa protein abundant in the beans.…”

Contact Urticaria to Raw Potato and Lentil Anaphylaxis: A Case Report

“…Kuntz-type soybean trypsin inhibitors can be isolated from legumes. 21 In our case, both contact urticaria with raw potatoes and lentil-induced anaphylaxis may be explained by Kunitz-type soybean trypsin inhibitors. Potatoes and legumes (lentil) contain this cross-reactive allergen protein.…”

Purification and characterization of a protein with antifungal, antiproliferative, and HIV‐1 reverse transcriptase inhibitory activities from small brown‐eyed cowpea seeds

“…Antifungal proteins from Nepalese red bean and cloud bean have antiproliferative activity but no anti‐HIV RT activity . The trypsin inhibitor from Gymnocladus sinensis has antiproliferative activity but not anti‐HIV RT activity , whereas ribonuclease from Schizophyllum commune has anti‐HIV RT activity but no antiproliferative activity . The trypsin inhibitor from white cloud bean has neither activity .…”