TcTI, a Kunitz-type trypsin inhibitor from cocoa associated with defense against pathogens

Protease inhibitors (PIs) are important biotechnological tools of interest in agriculture. Usually they are the first proteins to be activated in plant-induced resistance against pathogens. Therefore, the aim of this study was to characterize a Theobroma cacao trypsin inhibitor called TcTI. The ORF has 740 bp encoding a protein with 219 amino acids, molecular weight of approximately 23 kDa. rTcTI was expressed in the soluble fraction of Escherichia coli strain Rosetta [DE3]. The purified His-Tag rTcTI showed inhibitory activity against commercial porcine trypsin. The kinetic model demonstrated that rTcTI is a competitive inhibitor, with a Ki value of 4.08 × 10–7 mol L−1. The thermostability analysis of rTcTI showed that 100% inhibitory activity was retained up to 60 °C and that at 70–80 °C, inhibitory activity remained above 50%. Circular dichroism analysis indicated that the protein is rich in loop structures and β-conformations. Furthermore, in vivo assays against Helicoverpa armigera larvae were also performed with rTcTI in 0.1 mg mL−1 spray solutions on leaf surfaces, which reduced larval growth by 70% compared to the control treatment. Trials with cocoa plants infected with Mp showed a greater accumulation of TcTI in resistant varieties of T. cacao, so this regulation may be associated with different isoforms of TcTI. This inhibitor has biochemical characteristics suitable for biotechnological applications as well as in resistance studies of T. cacao and other crops.

Re-Print-Anaphylaxis Induced by Beer

The prevalence of cereal allergy is highly influenced by geographical area and consumption habits. According to data from Alergologica 2015 [1], cereal allergy accounts for 2.1% of all cases of food allergy in Spain. Beer is a barley-based alcoholic beverage that also contains hops, yeast, and other cereals (wheat, oats, corn, and even rye). Barley is also the basis of malt vinegar, whiskey, and gin. Several allergens of barley have been described, including lipid transfer protein (LTP), - and ß-amylase, gliadin, glutenin, peroxiredoxin, thionin, and trypsin inhibitor [2].

In-vitro and In-vivo effects of Cu-Melo Seed´trypsinseed´trypsin Inhibitor on Angiogenesis and Tumor Characteristics in Balb-c Mice With Breast Cancer

BackgroundMelon seeds as an excellent supply of protease inhibitors may have a protective role against tumor progression and angiogenesis. However, its effects on angiogenesis and the mechanism of its motion on cancer progression remain elusive. This study aimed to investigate the effect of bioactive compounds of melon seed on the expression of angiogenesis genes in breast cancer cell lines.MethodsTrypsin inhibitor (TI) was purified from the seed powder of Cucumis Melo. Half-maximal inhibitory concentration (IC50) was determined for TI, extract of melon seed powder (EXT), and tamoxifen (TAM) by MTT test. Also, breast tumor was induced by subcutaneous injection of MC4-L2 cell line in blab-c inbreed mice breast tissue. After tumor growth, mice were treated with TI, EXT, and TAM to examine their effects on the tumor characteristics and the expression of the angiogenesis-related genes including MMP-2, MMP-9, and VEGF using the RT-PCR method.ResultsTI, EXT, TAM, and adjuvant treatment of TI+TAM resulted a reduction in expression of MMP-2, MMP-9, and VEGF. All treatments improved the breast tumor characteristics and the necrosis. The Real Time-PCR method verified the positive effects of the treatments on the breast cancer cell line and tumors.ConclusionThe results indicated that treatments with trypsin Inhibitor Purified from Cucumis Melo Seeds and also combination therapy of trypsin inhibitor and tamoxifen can be considered as an alternative therapy in breast cancer patients. Further studies are warranted.

Small Molecule Peptidomimetic Trypsin Inhibitors: Validation Of An EKO Binding Mode, But With A Twist

Examination of a series of naturally-occurring trypsin inhibitor proteins, led to identification of a set of three residues (which we call the “interface triplet”) to be determinant of trypsin binding...

A novel anti-inflammatory activity of mcIRBP from Momordica charantia is associated with the improvement of diabetic nephropathy

Diabetic nephropathy is an inflammatory immune disorder accompanying diabetes. mcIRBP, the trypsin inhibitor of Momordica charantia, is an abundant 68-amino-acid-residue protein that interacts with the insulin receptor. Here the long-term...