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. 2017 Jan 23;22(1):187.
doi: 10.3390/molecules22010187.

Isolation and Characterization of a Phaseolus vulgaris Trypsin Inhibitor with Antiproliferative Activity on Leukemia and Lymphoma Cells

Miao Li  1   2 , Qin Liu  3 , Yajuan Cui  4 , Dong Li  5 , Hexiang Wang  6 , Tzi Bun Ng  7
Affiliations

Isolation and Characterization of a Phaseolus vulgaris Trypsin Inhibitor with Antiproliferative Activity on Leukemia and Lymphoma Cells

Miao Li et al. Molecules. .

Abstract

A 17.5-kDa trypsin inhibitor was purified from Phaseolus vulgaris cv. "gold bean" with an isolation protocol including ion exchange chromatography on DEAE-cellulose (Diethylaminoethyl-cellulose), affinity chromatography on Affi-gel blue gel, ion exchange chromatography on SP-sepharose (Sulfopropyl-sepharose), and gel filtration by FPLC (Fast protein liquid chromatography) on Superdex 75. It dose-dependently inhibited trypsin with an IC50 value of 0.4 μM, and this activity was reduced in the presence of dithiothreitol in a dose- and time-dependent manner, signifying the importance of the disulfide linkage to the activity. It inhibited [methyl-³H] thymidine incorporation by leukemia L1210 cells and lymphoma MBL2 cells with an IC50 value of 2.3 μM and 2.5 μM, respectively. The inhibitor had no effect on fungal growth and the activities of various viral enzymes when tested up to 100 μM.

Keywords: leguminous seeds; leukemia L1210; lymphoma MBL2; purification.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
SDS-PAGE results of gold bean trypsin inhibitor (GBTI, which was fraction SU2, 12 μg) and molecular mass marker proteins from GE Healthcare. Molecular mass of trypsin inhibitor was 17.5 kDa. The marker proteins included phosphorylase b (94 kDa), bovine serum albumin (67 kDa), ovalbumin (43 kDa), carbonic anhydrase (30 kDa), soybean trypsin inhibitor (20 kDa), and α-lactalbumin (14.4 kDa).
Figure 2
Figure 2
Inhibition of trypsin by GBTI. IC50 = 0.4 μM. Results are presented as mean ± SD (n = 3).
See this image and copyright information in PMC

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