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651.
Hiba Shabir Swati Kundu Seemi Farhat Basir Luqman A. Khan 《Biological trace element research》2014,160(1):116-122
Exposure to lead is known to cause vasoconstriction, exact mechanism of which remains to be elucidated. In this study, we investigate contractile responses of rat aortal rings equilibrated with Pb(II) in organ bath system, explore pathways responsible for hypercontraction and examine two ameliorators of lead-induced hypercontraction. At 1 μmol L?1 Pb(II), aortal rings showed an average increase of 50 % in isometric contraction. Incubation of rings, unexposed to Pb(II), with 1 μmol L?1 sodium nitroprusside (nitric oxide (NO) donor), 100 μmol L?1 apocynin (reactive oxygen species (ROS) inhibitor), and 100 μmol L?1 indomethacin (cyclooxygenase inhibitor) lead to decrease in phenylephrine-induced contraction by 31, 27, and 29 %, respectively. This decrease of contraction for Pb(II)-exposed rings was 48, 53, and 38 %, respectively, indicating that ROS- and NO-dependent components of contractions are significantly elevated in Pb(II)-induced hypercontraction. Cyclooxygenase-dependent contractile component did not show significant elevation. Eugenol and carvacrol are plant-derived phenols known to possess antioxidant activity and hence could act as possible ameliorators of hypercontraction. At saturating concentrations of 100 μmol L?1, eugenol and carvacrol caused a decrease in contraction by 38 and 42 % in unexposed rings and 46 and 50 % in Pb(II)-exposed rings. Co-incubation of rings with eugenol/carvacrol and various inhibitors suggests that both these active principles exert their relaxant effect via quenching of ROS and stimulation of NO synthesis. To conclude, Pb(II) is shown to induce hypercontraction of aortal rings through elevation of ROS and depletion of NO. This hypercontraction is effectively mitigated by eugenol and carvacrol. 相似文献
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Sheetal Uppal Shikha Salhotra Nitika Mukhi Fatima Kamal Zaidi Manas Seal Somdatta Ghosh Dey Rajiv Bhat Suman Kundu 《The Journal of biological chemistry》2015,290(4):1979-1993
Heme proteins, which reversibly bind oxygen and display a particular fold originally identified in myoglobin (Mb), characterize the “hemoglobin (Hb) superfamily.” The long known and widely investigated Hb superfamily, however, has been enriched by the discovery and investigation of new classes and members. Truncated Hbs typify such novel classes and exhibit a distinct two-on-two α-helical fold. The truncated Hb from the freshwater cyanobacterium Synechocystis exhibits hexacoordinate heme chemistry and bears an unusual covalent bond between the nonaxial His117 and a heme porphyrin 2-vinyl atom, which remains tightly associated with the globin unlike any other. It seems to be the most stable Hb known to date, and His117 is the dominant force holding the heme. Mutations of amino acid residues in the vicinity did not influence this covalent linkage. Introduction of a nonaxial His into sperm whale Mb at the topologically equivalent position and in close proximity to vinyl group significantly increased the heme stability of this prototype globin. Reversed phase chromatography, electrospray ionization-MS, and MALDI-TOF analyses confirmed the presence of covalent linkage in Mb I107H. The Mb mutant with the engineered covalent linkage was stable to denaturants and exhibited ligand binding and auto-oxidation rates similar to the wild type protein. This indeed is a novel finding and provides a new perspective to the evolution of Hbs. The successful attempt at engineering heme stability holds promise for the production of stable Hb-based blood substitute. 相似文献
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Plant Growth Regulation - Seed germination is one of the most crucial and complex physiological phenomena in the lifecycle of a plant, which often falls prey to environmental and biological stress... 相似文献
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Sheetal Uppal Amit Kumar Manish Shandilya Nitika Mukhi Amit Kumar Singh Suneel Kateriya Jagreet Kaur Suman Kundu 《Analytical biochemistry》2016
Hemoglobins with diverse characteristics have been identified in all kingdoms of life. Their ubiquitous presence indicates that these proteins play important roles in physiology, though function for all hemoglobins are not yet established with certainty. Their physiological role may depend on their ability to bind ligands, which in turn is dictated by their heme chemistry. However, we have an incomplete understanding of the mechanism of ligand binding for these newly discovered hemoglobins and the measurement of their kinetic parameters depend on their coordination at the heme iron. To gain insights into their functional role, it is important to categorize the new hemoglobins into either penta- or hexa-coordinated varieties. We demonstrate that simple pH titration and absorbance measurements can determine the coordination state of heme iron atom in ferric hemoglobins, thus providing unambiguous information about the classification of new globins. This method is rapid, sensitive and requires low concentration of protein. Penta- and hexa-coordinate hemoglobins displayed distinct pH titration profiles as observed in a variety of hemoglobins. The pentacoordinate distal histidine mutant proteins of hexacoordinate hemoglobins and ligand-bound hexacoordinate forms of pentacoordinate hemoglobins reverse the pH titration profiles, thus validating the sensitivity of this spectroscopic technique. 相似文献
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Jayasha Shandilya Parijat Senapati Karthigeyan Dhanasekaran Suma S. Bangalore Manoj Kumar A. Hari Kishore Akshay Bhat Gopinath S. Kodaganur Tapas K. Kundu 《FEBS letters》2014
The functional association of NPM1 with Aurora kinases is well documented. Surprisingly, although NPM1 is a well characterized phosphoprotein, it is unknown whether it is a substrate of Aurora kinases. We have found that Aurora kinases A and B can phosphorylate NPM1 at a single serine residue, Ser125, in vitro and in vivo. Phosphorylated-S125-NPM1 (pS125-NPM1) localizes to the midbody region during late cytokinesis where it colocalizes with Aurora B. The overexpression of mutant (S125A) NPM1 resulted in the deregulation of centrosome duplication and mitotic defects possibly due to cytokinesis failure. These data suggest that Aurora kinase B-mediated phosphorylation of NPM1 plays a critical role during mitosis, which could have wider implications in oncogenesis. 相似文献
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Predicting protein folding rate change upon point mutation using residue‐level coevolutionary information 下载免费PDF全文
Change in folding kinetics of globular proteins upon point mutation is crucial to a wide spectrum of biological research, such as protein misfolding, toxicity, and aggregations. Here we seek to address whether residue‐level coevolutionary information of globular proteins can be informative to folding rate changes upon point mutations. Generating residue‐level coevolutionary networks of globular proteins, we analyze three parameters: relative coevolution order (rCEO), network density (ND), and characteristic path length (CPL). A point mutation is considered to be equivalent to a node deletion of this network and respective percentage changes in rCEO, ND, CPL are found linearly correlated (0.84, 0.73, and ?0.61, respectively) with experimental folding rate changes. The three parameters predict the folding rate change upon a point mutation with 0.031, 0.045, and 0.059 standard errors, respectively. Proteins 2016; 84:3–8. © 2015 Wiley Periodicals, Inc. 相似文献