ABSTRACT
Calpain 10 plays a role in insulin secretion, action and susceptibility to type 2 diabetes. The mechanism through which it influences the insulin secretion and action is not completely defined. A structural bioinformatics approach is applied to envision its mechanism of action using available tools on NCBI (blastp and blastn), EMBL-EBI, Ensembl, Swiss Model Repository websites, I-TASSER, PROCHECK program and Discovery Studio software. Homology of domain I and II of calpain10 (isoform a) was established with super family cysteine proteinase domains (II a and II b, e=1.30e-77, 1.00e-20). Remaining sequences of domain III and T from (isoform a and c) indicated some similarity (Avg. e=1.94e-37) to calpain large subunit domain III (PF01067), the isoform g (139 AA) showed similarity with a part of catalytic domain of cysteine protease super family (e-value 1.00e-20). Swiss-model repository for 3D structures of protein, showed structural resemblance of 29% with 1QXP template of mu-calpain, 27% with 1KFX of m-calpain and 32% with 2P0R of calpain 9 in complex with leupeptin. Models prepared through I-TASSER confirmed through Ramachandran (RC) plots. The calpain 10 isoforms a, c and g show partial structural and functional resemblance to m, mu and calpain 9. This information is useful to find new drugs for disease management.
Subject(s)
Calpain/chemistry , Calpain/metabolism , Models, Molecular , Calpain/genetics , Computational Biology/methods , Humans , Protein Conformation , Protein Domains , Protein Isoforms/chemistry , Protein Isoforms/genetics , Protein Isoforms/metabolism , Sequence AlignmentABSTRACT
We aimed to investigate the effects of herbal St. John's Wort (SJW) on transcriptional regulation of hepatic tryptophan 2, 3 - dioxygenase (TDO) enzyme activity and brain regional serotonin (5-HT) levels in rats exposed to forced swim test (FST). TDO mRNA expression was quantified using real-time reverse transcription polymerase chain (RT-PCR) reaction and brain regional indoleamines were determined by high performance liquid chromatography coupled to fluorescence detector. Behavioral analysis shows significant reduction in immobility time in SJW (500mg/kg/ml) administered rats. It was found that pretreatment of SJW to rats did not prevent stress-induced elevation in plasma corticosterone levels however it increases serotonin synthesis by virtue of inhibiting hepatic TDO enzyme activity and its gene expression, ascertaining the notion that there exists an inverse relationship between hepatic TDO enzyme activity and brain 5-HT. The drug also decreases serotonin turnover in all the brain areas (hypothalamus, hippocampus amygdala) in stressed rats endorsing its monoamine oxidase inhibition property. Inhibition of TDO enzyme activity and its gene expression by the drug provides new insights for the development of therapeutic interventions for stress related mental illnesses.
Subject(s)
Antidepressive Agents/pharmacology , Brain/drug effects , Enzyme Inhibitors/pharmacology , Hypericum , Liver/drug effects , Plant Extracts/pharmacology , Serotonin/biosynthesis , Stress, Psychological/drug therapy , Tryptophan Oxygenase/antagonists & inhibitors , Animals , Behavior, Animal/drug effects , Brain/metabolism , Corticosterone/blood , Disease Models, Animal , Gene Expression Regulation, Enzymologic/drug effects , Liver/enzymology , Male , Motor Activity/drug effects , Phytotherapy , Plants, Medicinal , RNA, Messenger/metabolism , Rats , Rats, Wistar , Stress, Psychological/enzymology , Stress, Psychological/genetics , Stress, Psychological/psychology , Transcription, Genetic/drug effects , Tryptophan Oxygenase/genetics , Tryptophan Oxygenase/metabolism , Up-RegulationABSTRACT
Hyperglycemia associated with type 1 diabetes (T1D) alters the host immune system, resulting in a predisposition to infectious diseases. The high risk of infection in the diabetic population may lead to life-threatening situations. The early proteins of the alternative complement system pathway, constituting factors P, B, and D, have been shown to play an important role in preventing infection because they form a membrane attack complex (MAC)-C5-9, which debilitates the target microbes and/or molecules via cytotoxic and cytolytic reactions. Patients who are devoid of or contain low levels of these proteins may be susceptible to developing chronic infections. We have observed striking differences in partially fractionated serum proteins in diabetic patients (type 2) relative to controls, through single and two-dimensional gel electrophoresis. Our data, obtained from 50 diabetic patients in the age group of 25-45 years, who had the disease for fewer than 5 years, indicated patterns in low- and high-molecular-weight proteins, which could be grouped into five different categories with minor differences in their respective levels of protein expression. Immunoblot assay could barely detect the presence of properdin expression in diabetic patients. Quantization by ELISA in 99 patients indicated low levels of properdin expression in 70% of 50 diabetic patients (6.5 +/- 3 mug/mL) when compared to nondiabetic controls (19.5 +/- 8.5 mug/mL). This study concluded that patients with low expression of properdin should be advised to take extensive preventive measures and seek early management with appropriate treatments against infection.
