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The aim of this study was to investigate the potential of quercetin and two of its “in vivo“ metabolites, 3′-O-methyl quercetin and 4′-O-methyl quercetin, to protect H9c2 cardiomyoblasts against H 2 O 2 -induced oxidative stress. As limited data are available regarding the potential uptake and cellular effects of quercetin and its metabolites in cardiac cells, we have evaluated the...
We present circular dichroism (CD), steady state fluorescence and multidimensional NMR investigations on the equilibrium unfolding of monomeric dynein light chain protein (DLC8) by urea and guanidine hydrochloride (GdnHCl). Quantitative analysis of the CD and fluorescence denaturation curves reveals that urea unfolding is a two-state process, whereas guanidine unfolding is more complex. NMR investigations...
The linear IgE-binding epitopes of non-specific lipid transfer proteins (nsLTP) from plants were predicted using a combination of predictive tools including (1) the hydropathic profiles based on different scales of hydrophilicity, flexibility and exposure to the solvent, (2) the hydrophobic cluster analysis plots, (3) the occurrence of charged residues in the predicted amino acid sequence stretches...
AAV1, an alkaline glycoprotein (GP), was purified from Agkistrodon acutus venom by two chromatographic steps on successive DEAE-Sephadex A-50 and Superdex 75 FPLC columns. AAV1 on SDS–PAGE under non-reducing conditions migrated as a monomeric and a polymeric forms with apparent molecular mass of 57 and 180kDa, respectively. Upon reduction, it appeared as a single broad band with a mass of 50.3kDa...
RNA editing in plant mitochondria alters the RNA sequence by converting C-to-U or U-to-C at a specific site. We investigated the requirement for RNA editing in the complete genomic sequences of the gene encoding the cytochrome oxidase subunit III (cox3) and the ribosomal protein S13 (rps13) in 59 closely related species within the Asparagales and Liliales (monocots). To obtain a comprehensive picture...
In a previous paper, we reported more efficient enterokinase cleavage at a C-terminal non-target LKGDR 201 site compared with an internally sited canonical recognition site, DDDDK 156 . When this non-target site was placed internally to replace DDDDK 156 between the thioredoxin moiety and mouse NT-proCNP(1–50), this site was poorly processed leading us to conclude that efficient...
A series of N,N′-bis(2-pyridinylmethyl)diamines was synthesized and characterized for their inhibition effects towards plant copper-containing amine oxidase (EC 1.4.3.6) and polyamine oxidase (EC 1.5.3.11), which mediate the catabolic regulation of cellular polyamines. Even though these enzymes catalyze related reactions and, among others, act upon two common substrates (spermidine and spermine),...
A novel fibrinolytic serine protease has been identified and purified to homogeneity from the coelomic fluid of polychaete Nereis (Neanthes) virens (Sars), and named N-V protease. N-V protease is a 29kDa single chain protein with an isoelectric point of pH 4.5. It hydrolyzes Aα-chain of fibrinogen with a high efficiency, and the Bβ- and γ-chains (Aα>Bβ>γ) with a lower efficiency. The proteolytic...
Endothelial nitric oxide synthase (eNOS) plays a crucial role in the regulation of a variety of cardiovascular functions. Many studies have shown that dietary n-3 polyunsaturated fatty acids (PUFAs) have beneficial effects on coronary atherosclerosis. However, the mechanisms of n-3 PUFAs regulation in eNOS activation remain unknown. In the present study we investigated the effects of eicosapentaenoic...
Rho-associated kinase (Rho-kinase/ROCK/ROK) is a serine/threonine kinase and plays an important role in various cellular functions. The cAMP-dependent protein kinase (protein kinase A/PKA) and protein kinase C (PKC) are also serine/threonine kinases, and directly and/or indirectly take part in the signal transduction pathways of Rho-kinase. They have similar phosphorylation site motifs, RXXS/T and...
Histone lysine methylation plays a fundamental role in chromatin organization and function. This epigenetic mark is involved in many biological processes such as heterochromatin formation, chromosome X inactivation, genomic imprinting and transcriptional regulation. Here, we review recent advances in how histone lysine methylation participates in these biological events, and the enzymes that control...
Polynucleotide phosphorylase (PNPase) is a phosphate-dependent 3′ to 5′ exonuclease widely diffused among bacteria and eukaryotes. The enzyme, a homotrimer, can also be found associated with the endonuclease RNase E and other proteins in a heteromultimeric complex, the RNA degradosome. PNPase negatively controls its own gene (pnp) expression by destabilizing pnp mRNA. A current model of autoregulation...
We successfully cloned and expressed a single-chain antibody (425scFv), that is directed to human epidermal growth factor receptor HER1 (EGFR) in transgenic tobacco plants as a fusion with bacterial barstar gene (425scFv-barstar). Plant-produced recombinant 425scFv-barstar was recovered using barstar–barnase system. Based on barstar–barnase affinity, during purification of the plant-produced 425scFv-barstar,...
17β-Hydroxysteroid dehydrogenase from the fungus Cochliobolus lunatus (17β-HSDcl) is an NADPH-dependent member of the short-chain dehydrogenase/ reductase superfamily. To study the catalytic properties of this enzyme, we prepared several specific mutations of 17β-HSDcl (Tyr167Phe, His164Trp/Gly, Tyr212Ala). Wild-type 17β-HSDcl and the 17β-HSDcl mutants were evaluated by chromatographic, kinetic and...
Two isoforms of α-glucosidases (ONG2-I and ONG2-II) were purified from dry rice seeds (Oryza sativa L., var Nipponbare). Both ONG2-I and ONG2-II were the gene products of ONG2 mRNA expressed in ripening seeds. Each enzyme consisted of two components of 6kDa-peptide and 88kDa-peptide encoded by this order in ONG2 cDNA (ong2), and generated by post-translational proteolysis. The 88kDa-peptide of ONG2-II...
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