ABSTRACT
BACKGROUND AND OBJECTIVE: Protease inhibitors (PIs) regulate various cellular processes like cell cycle, differentiation, apoptosis and immune responses. Leguminous seeds are rich sources of protease inhibitors and many novel protease inhibitors have been purified from them. To isolate and purify protease inhibitors from seeds of Sophora japonica, characterize and investigate their anti- microbial activity. MATERIALS AND METHODS: Protease inhibitors (SJ-pi I and SJ-pi II) were purified to homogeneity by ammonium sulfate precipitation, Ion exchange chromatography and column chromatography. The molecular mass was estimated by size exclusion chromatography and by SDS-PAGE and anti- microbial activity was tested by agar disk diffusion method. RESULTS: Two protease inhibitors were isolated and purified from Sophora japonica seeds, SJ-pi I and SJ-pi II, with molecular weight of 15.1 and 31 kDa, respectively. Both purified inhibitors were active over a range of pH (6.0-9.0) and showed maximum activity in the temperature range of 30-40°C. They inhibited the growth of three Gram-positive bacteria. CONCLUSION: Protease inhibitors were classified as serine protease inhibitors, however further necessary structural investigations need to be carried out so as to group them into specific class of serine protease inhibitors.
Subject(s)
Anti-Infective Agents/pharmacology , Seeds/chemistry , Serine Proteinase Inhibitors/pharmacology , Sophora/chemistry , Anti-Infective Agents/isolation & purification , Apoptosis , Cell Cycle , Cell Differentiation , Chromatography, Ion Exchange , Chymotrypsin/chemistry , Gram-Positive Bacteria , Hydrogen-Ion Concentration , Immune System , Molecular Weight , Phytochemicals/isolation & purification , Phytochemicals/pharmacology , Plant Proteins/chemistry , Serine Proteinase Inhibitors/isolation & purification , TemperatureABSTRACT
Breast carcinogenesis is a multistep process, involving both genetic and epigenetic modification process of genes, involved in diverse pathways ranging from DNA repair to metabolic processes. This study was undertaken to assess the role of promoter methylation of GSTP1 gene, a member of glutathione-S-transferase family of enzymes, in relation to its expression, polymorphism, and clinicopathological parameters. Tissue samples were taken from breast cancer patients and paired with their normal adjacent tissues. A total of 51 subjects were studied, in which the frequency of promoter methylation in cancerous tissue was 37.25% as against 11% in the normal tissues ( p ≤ 0.001). The hypermethylated status of the gene was significantly associated with the loss of the protein expression ( r = -0.449, p = 0.001, odds ratio = 7.42, 95% confidence interval = 2.05-26.92). Furthermore, when compared with the clinical parameters, the significant association was found between the promoter hypermethylation and lymph node metastasis ( p ≤ 0.001), tumor stage ( p = 0.039), tumor grade ( p = 0.028), estrogen receptor status ( p = 0.018), and progesterone receptor status ( p = 0.046). Our study is the first of its kind in Kashmiri population, which indicates that GSTP1 shows aberrant methylation pattern in the breast cancer with the consequent loss in the protein expression. Furthermore, it also shows that the gene polymorphism (Ile105Val) at codon 105 is not related to the promoter methylation and two are the independent events in breast cancer development.
