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701.
BackgroundCathepsin H (E.C.3.4.22.16) belongs to a family of lysosomal cysteine protease which regulates diverse normal biological processes mainly in intracellular proteolysis.MethodsPurification of cathepsin H from an unstudied system i.e. buffalo lung has been achieved by a simple process developed after incorporating appropriate alteration in the available methods for isolation of the enzyme from other sources. The use of DEAE-Cellulose and SP-Sephadex C-50 helped in better and simultaneous separation of cathepsin B and H up to homogeneity.ResultsThe SDS-PAGE result showed buffalo cathepsin H to be a single-chain molecule having MW, NH2- and COOH- terminal residues of 25.4 kDa, Lys and Val respectively. The enzyme was a glycoprotein with pI of 6.2; it hydrolyzed Leu-NA (Vmax/Km = 301.6) as the most efficient substrate followed by Arg-NA, Arg-Arg-NA and BANA. Buffalo enzyme showed maximum activity at 36 °C, pH 6.75 and at a buffer concentration of 2 × 10?3 M.ConclusionCatheptic activity was found to be quite stable at least for 20–30 min between pH 4.5–7.0, buffer concentration of 1 × 10?2 to 4 × 10?2 M and the temperature resistance up to 36 °C. The effects of various substances present in the buffers routinely used for the assay of catheptic activity revealed that the activity of buffalo lung cathepsin H depends not only qualitatively but also quantitatively on the constituents of assay buffer.General significanceThis study seems to provide valuable information regarding the biochemistry of cathepsin H in general as well as influence of buffer constituents on enzyme activity and physiological role in particular. 相似文献
702.
703.
S Nair A Joshi-Saha S Singh R V S Singh V Thorat CP Kaushik S Eapen SF D'Souza 《Journal of biotechnology》2012,161(4):422-428
Phytoremediation is a viable strategy for management of toxic wastes in a large area/volume with low concentrations of toxic elemental pollutants. With increased industrial use of cobalt and its alloys, it has become a major metal contaminant in soils and water bodies surrounding these industries and mining sites with adverse effects on the biota. A bacterial Co-Ni permease was cloned from Rhodopseudomonas palustris and introduced into Nicotiana tabacum to explore its potential for phytoremediation and was found to be specific for cobalt and nickel. The transgenic plants accumulated more cobalt and nickel as compared to control, whereas no significant difference in accumulation of other divalent ions was observed. The transgenic plants were evaluated for cobalt content and showed increased acquisition of cobalt (up to 5 times) as compared to control. The plants were also assessed for accumulation of nickel and found to accumulate up to 2 times more nickel than control. At the same initial concentration of cobalt and nickel, transgenic plant preferentially accumulated cobalt as compared to nickel. The present study is perhaps the first attempt to develop transgenic plants expressing heterologous Co transporter with an improved capacity to uptake cobalt. 相似文献
704.
Pancreatic β-cell dysfunction is an early hallmark of type 1 diabetes mellitus. Among the potentially critical factors that cause β-cell dysfunction are cytokine attack, glucotoxicity, induction of endoplasmic reticulum (ER) or mitochondria stress. However, the exact molecular mechanism underlying β-cell's inability to maintain glucose homeostasis under severe stresses is unknown. This study used proinflammatory cytokines, thapsigargin, and rotenone in the presence of high concentration glucose to mimicking the conditions experienced by dysfunctional β-cells in human pancreatic islets, and profiled the alterations to the islet proteome with TMT-based proteomics. The results were further verified with label-free quantitative proteomics. The differentially expressed proteins under stress conditions reveal that immune related pathways are mostly perturbed by cytokines, while the respiratory electron transport chains and protein processing in ER pathways by rotenone. Thapsigargin together with high glucose induces dramatic increases of proteins in lipid synthesis and peroxisomal protein import pathways, with energy metabolism and vesicle secretion related pathways downregulated. High concentration glucose, on the other hand, alleviated complex I inhibition induced by rotenone. Our results contribute to a more comprehensive understanding of the molecular events involved in β-cell dysfunction. 相似文献
705.
Bottlegourd seedlings raised from antagonists coated seeds did not show any wilting symptoms and remained completely healthy.
The pathogen quantum inside the host tissue was found least. The pathogen propagules per gram of host tissue were restricted
and the soil population of theFusarium oxysporum was reduced very much in antagonists introduced soil. 相似文献
706.
O. Ramaswamy I. M. Saxena Sipra-Guha Mukherjee Sudhir K. Sopory 《Journal of biosciences》1983,5(1):63-69
In excised wheat leaves, the activity of nitrate reductase was enhanced by a brief pulse of red light and this increase was
reversed by far-red light irradiation. Even under continuous far-red light, nitrate reductase activity increased by 258% after
18 h. When leaves were kept in distilled water during exposure to red light and then transferred to potassium nitrate, there
was no difference in endogenous nitrate concentration. The nitrate reductase activity was the same whether leaves were floated
in potassium nitrate or in distilled water during irradiation. Partial to complete inhibition of enzyme activity was observed
when leaves were incubated in actinomycin-D and cycloheximide respectively, following 4 h of red light irradiation.In vitro irradiation of extract had no significant effect on nitrate reductase activity 相似文献
707.
Suman Kanji Ripon Sarkar Asmita Pramanik Sudhir Kshirsagar Carl J. Greene Hiranmoy Das 《Journal of cellular and molecular medicine》2021,25(5):2390-2403
Osteoclasts (OCs) differentiate from the monocyte/macrophage lineage, critically regulate bone resorption and remodelling in both homeostasis and pathology. Various immune and non-immune cells help initiating activation of myeloid cells for differentiation, whereas hyper-activation leads to pathogenesis, and mechanisms are yet to be completely understood. Herein, we show the efficacy of dental pulp–derived stem cells (DPSCs) in limiting RAW 264.7 cell differentiation and underlying molecular mechanism, which has the potential for future therapeutic application in bone-related disorders. We found that DPSCs inhibit induced OC differentiation of RAW 264.7 cells when co-cultured in a contact-free system. DPSCs reduced expression of key OC markers, such as NFATc1, cathepsin K, TRAP, RANK and MMP-9 assessed by quantitative RT-PCR, Western blotting and immunofluorescence detection methods. Furthermore, quantitative RT-PCR analysis revealed that DPSCs mediated M2 polarization of RAW 264.7 cells. To define molecular mechanisms, we found that osteoprotegerin (OPG), an OC inhibitory factor, was up-regulated in RAW 264.7 cells in the presence of DPSCs. Moreover, DPSCs also constitutively secrete OPG that contributed in limiting OC differentiation. Finally, the addition of recombinant OPG inhibited OC differentiation in a dose-dependent manner by reducing the expression of OC differentiation markers, NFATc1, cathepsin K, TRAP, RANK and MMP9 in RAW 264.7 cells. RNAKL and M-CSF phosphorylate AKT and activate PI3K-AKT signalling pathway during osteoclast differentiation. We further confirmed that OPG-mediated inhibition of the downstream activation of PI3K-AKT signalling pathway was similar to the DPSC co-culture–mediated inhibition of OC differentiation. This study provides novel evidence of DPSC-mediated inhibition of osteoclastogenesis mechanisms. 相似文献
708.
Ecosystems - Termites are dominant soil macro-fauna in tropical forests, yet remain an uncertain component of the carbon budget in many regions. We investigated CH4 and CO2 flux patterns from... 相似文献
709.
Syntheses are reported of 4-deoxy-d-xylo-hexose and 4-azido-4-deoxy-d-glucose as potential inhibitors for lactose synthase [uridine 5′-(α-d-galactopyranosyl pyrophosphate):d-glucose 4-β-d-galactopyranosyltransferase, EC 2.4.1.22]. These syntheses involved SN2 displacement of the 4-methylsulfonyloxy group of methyl 2,3,6-tri-O-benzoyl-4-O-methylsulfonyl-α-d-galactopyranoside by iodide and azide ions. In both cases, inversion in configuration was observed. The resulting intermediates, methyl 2,3,6-tri-O-benzoyl-deoxy-4-iodo-α-d-glucopyranoside and methyl 4-azido-2,3,6-tri-O-benzoyl-deoxy-α-d-glucopyranoside, were obtained in crystalline form. Both 4-deoxy-d-xylo-hexose and 4-azido-4-deoxy-d-glucose were found to be inhibitors for lactose synthase in the presence of α-lactalbumin, but had no effect in the absence of α-lactalbumin. Both d-glucose analogues bind to the enzyme system far more weakly than d-glucose, suggesting that the recognition of the 4-OH group of the acceptor substrate is an important factor in binding. 相似文献
710.