Trypsin Isoinhibitors with Antiproliferative Activity toward Leukemia Cells from<i>Phaseolus vulgaris</i>cv “White Cloud Bean”

Jian, Sun; Wang, Hexiang; Ng, Tzi Bun

doi:10.1155/2010/219793

Cited by 8 publications

(7 citation statements)

References 67 publications

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“…Otherwise, the inhibition mechanism of EvTI is different of a Kunitz-type inhibitor PdKI2 from Pithecelobium dumosum seed, which inhibits trypsin by competitive mode [63]. The IC 50 of EvTI for trypsin was 2.2×10 −8 mol.L −1 (0.42 µg.mL −1 ), that was less potent than urinary trypsin inhibitor (IC 50 of 6.44×10 −10 mol.L −1 ) [67], and lower than trypsin inhibitor from Sapindus saponaria seeds L (SSTI2 IC 50 8.3×10 −8 mol.L −1 ), and than purified trypsin inhibitors from Phaseolus vulgaris (IC 50 = 6.0×10 −7 mol.L −1 ) and from Glycine max (IC 50 = 1.9×10 −5 mol.L −1 ) [68], [69]. These results indicate that EvTI is effective on inhibiting trypsin like other inhibitors described in the literature.…”

Section: Discussionsupporting

confidence: 64%

Characterization and Pharmacological Properties of a Novel Multifunctional Kunitz Inhibitor from Erythrina velutina Seeds

et al. 2013

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Inhibitors of peptidases isolated from leguminous seeds have been studied for their pharmacological properties. The present study focused on purification, biochemical characterization and anti-inflammatory and anticoagulant evaluation of a novel Kunitz trypsin inhibitor from Erythrina velutina seeds (EvTI). Trypsin inhibitors were purified by ammonium sulfate (30–60%), fractionation followed by Trypsin-Sepharose affinity chromatography and reversed-phase high performance liquid chromatography. The purified inhibitor showed molecular mass of 19,210.48 Da. Furthermore, a second isoform with 19,228.16 Da was also observed. The inhibitor that showed highest trypsin specificity and enhanced recovery yield was named EvTI (P2) and was selected for further analysis. The EvTI peptide fragments, generated by trypsin and pepsin digestion, were further analyzed by MALDI-ToF-ToF mass spectrometry, allowing a partial primary structure elucidation. EvTI exhibited inhibitory activity against trypsin with IC50 of 2.2×10−8 mol.L−1 and constant inhibition (Ki) of 1.0×10−8 mol.L−1, by a non-competitive mechanism. In addition to inhibit the activity of trypsin, EvTI also inhibited factor Xa and neutrophil elastase, but do not inhibit thrombin, chymotrypsin or peptidase 3. EvTI was investigated for its anti-inflammatory and anti-coagulant properties. Firstly, EvTI showed no cytotoxic effect on human peripheral blood cells. Nevertheless, the inhibitor was able to prolong the clotting time in a dose-dependent manner by using in vitro and in vivo models. Due to anti-inflammatory and anticoagulant EvTI properties, two sepsis models were here challenged. EvTI inhibited leukocyte migration and specifically acted by inhibiting TNF-α release and stimulating IFN-α and IL-12 synthesis. The data presented clearly contribute to a better understanding of the use of Kunitz inhibitors in sepsis as a bioactive agent capable of interfering in blood coagulation and inflammation.

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Section: Discussionsupporting

confidence: 64%

Characterization and Pharmacological Properties of a Novel Multifunctional Kunitz Inhibitor from Erythrina velutina Seeds

et al. 2013

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“…From 100 g of small pinto beans, about 141 mg of trypsin inhibitor could be collected using the protocol mentioned above. The yield was higher than that obtained in purification of many other trypsin inhibitors, such as broad bean, white cloud bean and Faba bean trypsin inhibitors ($11, $29 and $54 mg/100 g beans, respectively) [22][23][24] .…”

Section: Discussionmentioning

confidence: 63%

“…The three-step protocol is simpler than purification procedures employed for many other trypsin inhibitors such as white cloud bean trypsin inhibitor 22 . By using fewer purification steps, the loss of the trypsin inhibitor can be minimized, and a higher yield of the protein can be achieved.…”

Section: Discussionmentioning

confidence: 99%

A thermostable trypsin inhibitor with antiproliferative activity from small pinto beans

Chan

Zhang

Sze

et al. 2013

Journal of Enzyme Inhibition and Medicinal Chemistry

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Small pinto bean is a cultivar of Phaseolus vulgaris. It produces a 16-kDa trypsin inhibitor that could be purified using anion exchange and size chromatography. Q-Sepharose, Mono Q and Superdex 75 columns were employed for the isolation process. Small pinto bean trypsin inhibitor demonstrated moderate pH stability (pH 2-10) and marked heat stability, with its trypsin inhibitory activity largely retained after exposure to 100 C for half an hour. The activity was abolished in the presence of dithiothreitol, in a dose-dependent manner, implying that disulfide bonds in small pinto bean trypsin inhibitor are crucial for the activity. The trypsin inhibitor showed a blocked N-terminus. The trypsin inhibitor only slightly inhibited the viability of breast cancer MCF7 and hepatoma HepG2 cells at 125 mM.

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“…The study also reported that cells treated with TBPI diminished their invasive ability most probably due to the suppression of MMP2 and MMP9. Sun et al (2010) isolated and purified two trypsin inhibitors of each with a molecular mass of 16 kDa from Phaseolus vulgaris cv “White Cloud Bean.” Both of these inhibitors showed an antiproliferative activity toward Leukemia L1210 albeit with a little variance in potency, but there was little activity toward lymphoma MBL2 cells.…”

Section: Roles Of Plant Protease Inhibitors In Health and Disease Conmentioning

confidence: 99%

Plant Protease Inhibitors in Therapeutics-Focus on Cancer Therapy

2016

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Plants are known to have many secondary metabolites and phytochemical compounds which are highly explored at biochemical and molecular genetics level and exploited enormously in the human health care sector. However, there are other less explored small molecular weight proteins, which inhibit proteases/proteinases. Plants are good sources of protease inhibitors (PIs) which protect them against diseases, insects, pests, and herbivores. In the past, proteinaceous PIs were considered primarily as protein-degrading enzymes. Nevertheless, this view has significantly changed and PIs are now treated as very important signaling molecules in many biological activities such as inflammation, apoptosis, blood clotting and hormone processing. In recent years, PIs have been examined extensively as therapeutic agents, primarily to deal with various human cancers. Interestingly, many plant-based PIs are also found to be effective against cardiovascular diseases, osteoporosis, inflammatory diseases and neurological disorders. Several plant PIs are under further evaluation in in vitro clinical trials. Among all types of PIs, Bowman-Birk inhibitors (BBI) have been studied extensively in the treatment of many diseases, especially in the field of cancer prevention. So far, crops such as beans, potatoes, barley, squash, millet, wheat, buckwheat, groundnut, chickpea, pigeonpea, corn, and pineapple have been identified as good sources of PIs. The PI content of such foods has a significant influence on human health disorders, particularly in the regions where people mostly depend on these kind of foods. These natural PIs vary in concentration, protease specificity, heat stability, and sometimes several PIs may be present in the same species or tissue. However, it is important to carry out individual studies to identify the potential effects of each PI on human health. PIs in plants make them incredible sources to determine novel PIs with specific pharmacological and therapeutic effects due to their peculiarity and superabundance.

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Trypsin Isoinhibitors with Antiproliferative Activity toward Leukemia Cells fromPhaseolus vulgariscv “White Cloud Bean”

Cited by 8 publications

References 67 publications

Characterization and Pharmacological Properties of a Novel Multifunctional Kunitz Inhibitor from Erythrina velutina Seeds

Characterization and Pharmacological Properties of a Novel Multifunctional Kunitz Inhibitor from Erythrina velutina Seeds

A thermostable trypsin inhibitor with antiproliferative activity from small pinto beans

Plant Protease Inhibitors in Therapeutics-Focus on Cancer Therapy

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