Measurement of Aspartate Aminotransferase-P2 in Developing Lupin Nodules Using a Two Site ELISA Employing Monoclonal Antibodies

1992 ◽  
Vol 19 (2) ◽  
pp. 147 ◽  
Author(s):  
WT Jones ◽  
SD Jones ◽  
CP Liddane ◽  
PHS Reynolds
Keyword(s):  
Monoclonal Antibodies ◽  
Catalytic Activity ◽  
Aspartate Aminotransferase ◽  
Root Nodule ◽  
Root Nodules ◽  
Lupinus Angustifolius ◽  
Elisa Method ◽  
Elisa System ◽  
Rhizobium Lupini ◽  
P2 Protein

Leguminous root nodules contain two forms of aspartate aminotransferase (AAT). AAT-P1 is con- stitutively expressed in roots and nodules, whereas the second isoenzyme, AAT-P2, is induced in developing nodules. A two site ELISA system was developed to quantify AAT-P2 in nodule extracts. The assay comprised two affinity-purified monoclonal antibodies directed against different epitopes on the AAT-P2 molecule. The sensitivity was less than 200 pg mL-1, and AAT-P2 could be quantified in the range 1 to 15 ng mL-1. The assay was specific for AAT-P2, and did not measure AAT-P1. The assay was used to specifically measure AAT-P2 in the developing legume root nodule formed by the symbiotic liaison between Rhizobium lupini NZP2257 and the roots of Lupinus angustifolius L. The ELISA method, which measures AAT-P2 protein, was contrasted with an approach which relied on the heat lability of AAT-P2 at 60°C to measure individual isoenzyme catalytic activity.

scholarly journals Production and Characterization of Monoclonal Antibodies against Aspartate Aminotransferase-P2 from Lupin Root Nodules

1990 ◽  
Vol 94 (3) ◽  
pp. 1358-1364 ◽  
Author(s):  
William T. Jones ◽  
Paul H. S. Reynolds ◽  
Stephen D. Jones ◽  
Cherry P. Liddane ◽  
Karen A. Rodber
Keyword(s):  
Monoclonal Antibodies ◽  
Aspartate Aminotransferase ◽  
Root Nodules

scholarly journals ELISA for Urinary Trehalase with Monoclonal Antibodies: A Technique for Assessment of Renal Tubular Damage

2000 ◽  
Vol 46 (5) ◽  
pp. 636-643 ◽  
Author(s):  
Reiko Ishihara ◽  
Shigeru Taketani ◽  
Misa Sasai-Takedatsu ◽  
Yasushi Adachi ◽  
Minoru Kino ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Monoclonal Antibodies ◽  
Catalytic Activity ◽  
Enzyme Activities ◽  
Proximal Tubules ◽  
Dent Disease ◽  
Renal Diseases ◽  
Tubular Damage ◽  
Renal Proximal Tubules ◽  
Elisa System

Abstract Background: α,α-Trehalase, located on renal proximal tubules, is a glycoprotein that hydrolyses α,α-trehalose to two glucose molecules. Urinary trehalase reflects damage to renal proximal tubules, but its activity has not been measured routinely because measurement of catalytic activity is rather complicated and because conventional assays for enzyme activity might not reflect all of the trehalase protein because of enzyme inactivation in urinary samples. Methods: We established novel monoclonal antibodies for human trehalase and a sandwich ELISA for quantification of urinary trehalase. We determined the urinary trehalase protein concentration with this ELISA and trehalase catalytic activity, and the results of these two methods were compared. Results: The ELISA system was more sensitive than the detection of enzyme activity and could detect a subtle difference in the amount of trehalase present in renal diseases. The within- and between-assay CVs in the ELISA were 6.7–7.6% and 6.2–8.2%, respectively. Highly significant increases in both the quantity and activity were seen in patients with nephrotic syndrome (acute phase), Lowe syndrome, and Dent disease. The quantities were 70- to 200-fold greater, whereas enzyme activities were, at most, 10-fold higher than those of control subjects. In the detection of small amounts of trehalase in patients with chronic glomerulonephritis and renal anomalies, quantities were better than enzyme activities. Conclusions: We have established an ELISA system for quantification of urinary trehalase that uses novel monoclonal antibodies. Our ELISA system is simpler and more sensitive than a conventional activity assay and reflects trehalase protein. This ELISA can be a useful as a common tool for clinical assessment of renal proximal tubular damage.

scholarly journals Production and Characterization of Monoclonal Antibodies against Aspartate Aminotransferase-P1 from Lupin Root Nodules

1994 ◽  
Vol 104 (1) ◽  
pp. 91-97 ◽  
Author(s):  
W. T. Jones ◽  
S. D. Jones ◽  
D. Harvey ◽  
K. R. Rodber ◽  
G. B. Ryan ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Aspartate Aminotransferase ◽  
Root Nodules

scholarly journals Aboveground plant-to-plant communication reduces root nodule symbiosis and soil nutrient concentrations

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yuta Takahashi ◽  
Kaori Shiojiri ◽  
Akira Yamawo
Keyword(s):  
Root Nodule ◽  
Total Phenolics ◽  
Root Nodules ◽  
Soil Nutrient ◽  
Chemical Defenses ◽  
Nutrient Concentrations ◽  
Soil C ◽  
Plant Communication ◽  
Root Nodule Symbiosis ◽  
Nodule Symbiosis

AbstractAboveground communication between plants is well known to change defense traits in leaves, but its effects on belowground plant traits and soil characteristics have not been elucidated. We hypothesized that aboveground plant-to-plant communication reduces root nodule symbiosis via induction of bactericidal chemical defense substances and changes the soil nutrient environment. Soybean plants were exposed to the volatile organic compounds (VOCs) from damaged shoots of Solidago canadensis var. scabra, and leaf defense traits (total phenolics, saponins), root saponins, and root nodule symbiosis traits (number and biomass of root nodules) were measured. Soil C/N ratios and mineral concentrations were also measured to estimate the effects of resource uptake by the plants. We found that total phenolics were not affected. However, plants that received VOCs had higher saponin concentrations in both leaves and roots, and fewer root nodules than untreated plants. Although the concentrations of soil minerals did not differ between treatments, soil C/N ratio was significantly higher in the soil of communicated plants. Thus, the aboveground plant-to-plant communication led to reductions in root nodule symbiosis and soil nutrient concentrations. Our results suggest that there are broader effects of induced chemical defenses in aboveground plant organs upon belowground microbial interactions and soil nutrients, and emphasize that plant response based on plant-to-plant communications are a bridge between above- and below-ground ecosystems.

scholarly journals Coalescence of rhizobial communities in soil interacts with fertilization and determines the assembly of rhizobia in root nodules

2020 ◽  
Author(s):  
Josep Ramoneda ◽  
Johannes Le Roux ◽  
Stefanie Stadelmann ◽  
Emmanuel Frossard ◽  
Beat Frey ◽  
...  
Keyword(s):  
Microbial Community ◽  
Soil Microbial Community ◽  
Root Nodule ◽  
Root Nodules ◽  
Arable Land ◽  
Soil Microbial ◽  
Soil Mixing ◽  
Environmental Setting ◽  
Widespread Phenomenon ◽  
The Individual

AbstractSoil microbial community coalescence, whereby entire microbial communities mix and compete in a new environmental setting, is a widespread phenomenon whose applicability for targeted root microbiome assembly has not been studied. Using a legume shrub adapted to nutrient poor soil, we tested for the first time how the assembly of communities of rhizobial root nodule symbionts is affected by the interaction of coalescence and fertilization. Seedlings of the rooibos [Aspalathus linearis (Burm.f.) Dahlg.], were raised in pairwise mixtures of soil from cultivated and uncultivated land of five farms, as well as the individual mixture components. A fragment of the symbiosis maker gene, nodA, was sequenced to characterize the taxonomic turnover of the rhizobia associated with all root nodules at the age of eight month. Soil mixing promoted taxonomic turnover in the rhizobial communities, while fertilization amplified such turnover by increasing the number of rhizobia that became more abundant after soil mixing. Soil mixing and fertilization had a synergistic effect on the abundance of a particular taxon, which was rare in the component soils but became highly abundant in fertilized plants raised in soil mixtures. These findings provide the first evidence that fertilizer addition can interact with soil microbial community coalescence, probably through increasing the chances for rare strains to prioritize root nodule colonization. The combination of soil mixing and fertilizer addition may be a still unexplored measure to (re)introduce root microbial mutualists in arable land.

Characterization of Monoclonal Antibodies that Inhibit the Catalytic Activity of Acetylcholinesterases

2002 ◽  
Vol 64 (2) ◽  
pp. 842-849 ◽  
Author(s):  
Mary K. Gentry ◽  
Deborah R. Moorad ◽  
Regina S. Hur ◽  
Ashima Saxena ◽  
Yacov Ashani ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Catalytic Activity

Rapid Detection of Salmonella spp. in Foods by Combination of a New Selective Enrichment and a Sandwich ELISA Using Two Monoclonal Antibodies against Dulcitol 1-Phosphate Dehydrogenase

1996 ◽  
Vol 59 (11) ◽  
pp. 1158-1163 ◽  
Author(s):  
HUAIZE TIAN ◽  
TAKAHISA MIYAMOTO ◽  
TAKASHI OKABE ◽  
YOICHIRO KURAMITSU ◽  
KEN-ICHI HONJOH ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Rapid Detection ◽  
Enrichment Culture ◽  
Sandwich Elisa ◽  
False Negative ◽  
Detection Sensitivity ◽  
Elisa Method ◽  
Salmonella Spp ◽  
Selective Enrichment ◽  
Raw Foods

A rapid-detection method was developed for food-borne dulcitol-positive Salmonella spp. in foods that involves a new preenrichment and selective enrichment system and a sandwich ELISA using two monoclonal antibodies against dulcitol 1-phosphate dehydrogenase. Preenrichment and selective enrichment were in Enterobacteriaceae enrichment mannitol (EEM) broth at 42°C for 6 h and in a new dulcitol-magnesium chloride-pyridinesulfonic acid brilliant green-novobiocin (DMPBN) medium at 42°C for 27 h, respectively. The cells were collected from the selective enrichment culture and suspended in 0.1 ml of 1 N NaOH for 2 min. The solution was neutralized with 0.1 ml of 2 M Tris-HCl buffer (pH 7.5) and the mixture was used as a sample for ELISA. The detection sensitivity of the ELISA was 105 CFU of Salmonella spp. per ml of culture. Competing non-Salmonella organisms in raw food did not interfere with the detection of Salmonella cells even when present at 107: 1 (non-Salmonella: Salmonella ratio) in food. Nonmotile Salmonella gallinarum was detected by the ELISA. The minimum detectable number of initial inoculum of Salmonella typhimurium was 0.69 CFU/25 g of raw chicken after the preenrichment in EEM broth and the selective enrichment in DMPBN medium. The present ELISA method required a total analysis time of 36 h including the preenrichment and selective enrichment periods. The ELISA method was compared with a conventional cultural method for the detection of Salmonella cells in 130 samples of raw foods. Of the samples tested, 16 were Salmonella-positive and 114 samples were negative by both methods. False-positive and false-negative results were not encountered.

Influence of substrate concentration on the activation of transketolase, aspartate aminotransferase, and glutathione reductase by coenzymes.

1984 ◽  
Vol 30 (1) ◽  
pp. 143-144
Author(s):  
J C Hafkenscheid ◽  
C M van Dijk
Keyword(s):  
Catalytic Activity ◽  
Glutathione Reductase ◽  
Aspartate Aminotransferase ◽  
Substrate Concentration ◽  
Activity Concentrations ◽  
Influence Of Substrate ◽  
Underlying Mechanisms ◽  
Substrate Concentrations

Abstract We investigated the mechanism by which the three most commonly measured enzymes in erythrocytes are activated by their respective coenzymes by determining the catalytic activity concentrations of transketolase (EC 2.2.1.1), aspartate aminotransferase (EC 2.6.1.1), and glutathione reductase (EC 1.6.4.2) in relation to various substrate concentrations. We conclude that the underlying mechanisms by which the enzymes are activated are not the same.

Identification of epitopes on ADAMTS13 recognized by a panel of monoclonal antibodies with functional or non-functional effects on catalytic activity

2012 ◽  
Vol 130 (3) ◽  
pp. e79-e83 ◽  
Author(s):  
Atsuko Igari ◽  
Terumichi Nakagawa ◽  
Takanori Moriki ◽  
Yusuke Yamaguchi ◽  
Masanori Matsumoto ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Catalytic Activity
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