Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:1.10.3.3 (ascorbate oxidase)
 778 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Equilibrium denaturation experiments have been performed in order to study the dissociation into monomers and unfolding of the dimeric copper-containing enzyme ascorbate oxidase by urea and guanidine hydrochloride. The process has been followed by fluorescence intensity and anisotropy, by optical density, and by circular dichroism as a function of denaturant concentration. The noncoincidence of the unfolding curves obtained by different techniques suggests that a multiphasic process is occurring. The study of enzymatic activity and aromatic circular dichroism as a function of denaturant concentration shows that the first transition involves a change in the protein tertiary structure which is accompanied by the loss of biological function. Gel electrophoresis, ultracentrifugation, and protein dilution experiments demonstrate that a large fraction of protein molecules is still dimeric during this first transition with a stability which is strictly dependent on the denaturant used. The free energy change from the native form to this intermediate species was estimated to be approximately 3.5 kcal/mol. The binding of 1-anilino-8-naphthalenesulfonic acid to the partially unfolded, inactive ascorbate oxidase dimer also suggests a large conformational change accompanied by copper release, allowing the probe to penetrate deep inside the protein structure. Further denaturation to give a fully unfolded form is protein concentration dependent, suggesting that dissociation into monomers is occurring. The monomers appear to be very unstable. No evidence for structured intermediates was in fact detected in the last step of the denaturation process. A three-state model has been used to fit the fluorescence data, and the fractions of different species have been calculated as a function of denaturant concentration. The total free energy change of the unfolding transition using either urea or guanidine hydrochloride is rather small ( approximately 15-16 kcal/mol) and quite comparable to the value found for smaller proteins. The loss of secondary structure which occurs in the second part of the unfolding transition may be described by a simple two-state process which is characterized by a free energy change of 12-13 kcal/mol. These results suggest that the folding process of ascorbate oxidase follows a hierarchical model (Jaenicke, 1991). In this context, the assembly of monomers in a dimeric molecule plays a fundamental role by enhancing the protein stability and driving the final organization of the tertiary structure.
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PMID:Role of quaternary structure in the stability of dimeric proteins: the case of ascorbate oxidase. 928 82

This experiment was conducted to determine the effect of crop processing and amino acid supplementation on dairy cow performance. Corn silage processed (PCS) or unprocessed (UCS) was used as the main forage (45% of dry matter, DM) in a total mixed ration (TMR). Each TMR was either supplemented (AA) or not (AAO) with ruminally protected amino acids (lysine, 3 g/d and methionine, 14 g/d). Thirty-two (551 kg) Holstein cows were randomly assigned to four treatments: PCS-AA, PCS-AA0, UCS-AA, and UCS-AA0 in a 2 x 2 factorial structure. Between wk 7 and 17 of lactation, cows were fed ad libitum TMR comprising 45% of corn silage plus 1 kg of grass hay once a day. The UCS presented better fermentation characteristics than PCS. Dry matter intake (DMI) of the TMR was not affected by treatment and averaged 22.7 kg/d. Energy-corrected milk (ECM) production was 9% higher with UCS than with PCS (33.1 vs. 30.1 kg/d). Milk efficiency was therefore 6% higher with UCS than with PCS (1.43 vs. 1.35 kg ECM/kg of DMI). The concentration of major milk constituents (fat, protein, lactose, urea) was not affected by treatments. Apparent digestibility of DM, organic matter, N, starch, acid detergent fiber, and neutral detergent fiber were similar among treatments. The effective ruminal degradability of DM, starch, and protein, however, was greater with PCS than with UCS. Amino acid supplementation had no effect on milk production nor on milk constituents, whether it was used with processed corn silage or with unprocessed corn silage. These data indicate that feeding UCS resulted in a greater milk production compared with PCS. The numerically higher DMI, a potentially greater intestinal digestion of starch or the better conservation of UCS could have contributed to the greater milk production.
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PMID:Effects of corn silage processing and amino acid supplementation on the performance of lactating dairy cows. 1467 98

Soil salinization is a serious problem for cultivation of rice, as among cereals rice is the most salt sensitive crop, and more than 40% of the total agricultural land amounting to approximately 80 million ha the world over is salt affected. Salinity affects a plant in a varieties of ways, including ion toxicity, osmotic stress and oxidative damage. Since miRNAs occupy the top place in biochemical events determining a trait, understanding their role in salt tolerance is highly desirable, which may allow introduction of the trait in the rice cultivars of choice through biotechnological interventions. High throughput sequencing of sRNAs in the root and shoot tissues of the seedlings of the control and NaCl treated Pokkali, a salt-tolerant rice variety, identified 75 conserved miRNAs and mapped 200 sRNAs to the rice genome as novel miRNAs. Expression of nine novel miRNAs and two conserved miRNAs were confirmed by Northern blotting. Several of both conserved and novel miRNAs that expressed differentially in root and/or shoot tissues targeted transcription factors like AP2/EREBP domain protein, ARF, NAC, MYB, NF-YA, HD-Zip III, TCP and SBP reported to be involved in salt tolerance or in abiotic stress tolerance in general. Most of the novel miRNAs expressed in the salt tolerant wild rice Oryza coarctata, suggesting conservation of miRNAs in taxonomically related species. One of the novel miRNAs, osa-miR12477, also targeted L-ascorbate oxidase (LAO), indicating build-up of oxidative stress in the plant upon salt treatment, which was confirmed by DAB staining. Thus, salt tolerance might involve miRNA-mediated regulation of 1) cellular abundance of the hormone signaling components like EREBP and ARF, 2) synthesis of abiotic stress related transcription factors, and 3) antioxidative component like LAO for mitigation of oxidative damage. The study clearly indicated importance of osa-miR12477 regulated expression of LAO in salt tolerance in the plant.
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PMID:Identification and expression analysis of miRNAs and elucidation of their role in salt tolerance in rice varieties susceptible and tolerant to salinity. 3229 92