Kinetics of Spanish broom peroxidase obeys a Ping-Pong Bi–Bi mechanism with competitive inhibition by substrates

Galende, Patricia Pérez; Cuadrado, Nazaret Hidalgo; Kostetsky, E. Ya.; Roig, Manuel G.; Villar, Enrique; Shnyrov, Valery L.; Kennedy, John F.

doi:10.1016/j.ijbiomac.2015.09.042

Cited by 28 publications

(23 citation statements)

References 33 publications

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“…Specifically, POD activity was increased at 16 (113.16%), 24 (114.56%), 72 (108.48%), 96 (131.94%), and 192 h (129.01%), but was decreased at 8 (90.68%), 48 (95.84), and 144 h (95.84%). These fluctuating kinetics has been described as a ping-pong bisubstrate mechanism for various PODs, including horseradish POD (Dunford, 1995;Deyhimi and Nami, 2011), tobacco anionic POD (Lagrimini et al, 1993) and Cistus multiflorus POD (Galende et al, 2015), and reflect the early release of water before the binding of an electron donor (Lagrimini et al, 1993;Deyhimi and Nami, 2011). These observations indicate that aleurone PODs could be subject to competitive substrate inhibition, as described above.…”

Section: Modulation Of Endosperm Pod Activity and Expression Of Zmprxmentioning

confidence: 72%

Modulation of Aleurone Peroxidases in Kernels of Insect-Resistant Maize (Zea mays L.; Pob84-C3R) After Mechanical and Insect Damage

et al. 2020

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Peroxidases (PODs) have many biological functions during the plant life cycle. In maize kernels, endosperm PODs have been identified as biochemical contributors to resistance against Sitophilus zeamais, but their identities have not been determined. In this study, we identified these PODs and determined whether their contributions are basal or inducible. Semi-purification and LC-MS/MS analyses showed that the protein ZmPrx35 is the predominant soluble endosperm POD from kernels of Pob84-C3R. Subsequent time-course analyses after mechanical damage showed that POD activity was regulated in a fluctuating kinetics pattern and that zmprx35 mRNA expression levels reflected this pattern. After 48 h of infestation with S. zeamais or Prostephanus truncatus, soluble endosperm POD activities were 1.38-or 0.85-fold, respectively. Under the same conditions, zmprx35 expression was induced 1.61-fold (S. zeamais infestation) and 1.17-fold (P. truncatus infestation). These findings suggest that ZmPrx35 contributes to the protective responses of aleurone cells against wounding and pest attacks, which could be enhanced/repressed by insect factors. Our data also provide evidence that the mechanisms of resistance of maize Pob84-C3R kernels toward the insect pests S. zeamais and P. truncatus are independent.

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Section: Modulation Of Endosperm Pod Activity and Expression Of Zmprxmentioning

confidence: 72%

Modulation of Aleurone Peroxidases in Kernels of Insect-Resistant Maize (Zea mays L.; Pob84-C3R) After Mechanical and Insect Damage

et al. 2020

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“…Previous mathematical models developed to describe kinetics of HRP on the electrodes [ 21 , 37 , 38 , 39 , 40 ] focused on the enzyme’s kinetics or were based on an assumption that the J is mass-transfer limited. In contrast, our model explicitly calculates the rates of all key reaction and mass transfer steps, all of which could limit the signal’s magnitude to some extent.…”

Section: Resultsmentioning

confidence: 99%

“…The non-linear, ping-pong kinetic mechanism describing HRP oxidation of C in the presence of

is shown in Reactions (i)–(iii) [ 33 , 34 , 35 , 36 ]: HRP (Fe 3+ ) + H 2 O 2 → Compound(I) + H 2 O Compound (I) + C → Compound (II) + Q Compound (II) + C → HRP (Fe 3+ ) + Q where compounds (I) and (II) are oxidized intermediates of HRP. The kinetic formula resulting from this mechanism [ 21 , 37 , 38 , 39 , 40 ] is:

where

is the reaction rate,

is the maximum reaction rate constant (

and [HRP] are turnover number and HRP concentration within the immunosensing layer, respectively;

and

are the corresponding Michaelis–Menten constants, and

and

are

and C concentrations, respectively.…”

Section: Mechanistic Mathematical Modelmentioning

confidence: 99%

Integrated Experimental and Theoretical Studies on an Electrochemical Immunosensor

et al. 2020

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Electrochemical immunosensors (EIs) integrate biorecognition molecules (e.g., antibodies) with redox enzymes (e.g., horseradish peroxidase) to combine the advantages of immunoassays (high sensitivity and selectivity) with those of electrochemical biosensors (quantitative electrical signal). However, the complex network of mass-transfer, catalysis, and electrochemical reaction steps that produce the electrical signal makes the design and optimization of EI systems challenging. This paper presents an integrated experimental and modeling framework to address this challenge. The framework includes (1) a mechanistic mathematical model that describes the rate of key mass-transfer and reaction steps; (2) a statistical-design-of-experiments study to optimize operating conditions and validate the mechanistic model; and (3) a novel dimensional analysis to assess the degree to which individual mass-transfer and reaction steps limit the EI’s signal amplitude and sensitivity. The validated mechanistic model was able to predict the effect of four independent variables (working electrode overpotential, pH, and concentrations of catechol and hydrogen peroxide) on the EI’s signal magnitude. The model was then used to calculate dimensionless groups, including Damkohler numbers, novel current-control coefficients, and sensitivity-control coefficients that indicated the extent to which the individual mass-transfer or reaction steps limited the EI’s signal amplitude and sensitivity.

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“…The reciprocal substrate concentration 1/[S] and the initial enzyme reaction rate 1/ v were plotted reciprocally. 20 According to the dynamic equation of the sequence mechanism, the following results: V = ( V max × [C][S])/([C][S] + [S] K c,m + [C] K s,m + K c,m K s,m ) where [C] means the concentration of coenzyme NADH and [S] is the concentration of substrate RB5. In a multi-substrate reaction, the Michaelis constant of one substrate changes with the concentration of the other substrate, where K c,m refers to the Michaelis constant of NADH if the concentration of dye RB5 reaches saturation.…”

Section: Methodsmentioning

confidence: 99%

Section: Kinetic Properties Of Bvu5mentioning

confidence: 99%

Biochemical characterization of a novel azo reductase named BVU5 from the bacterial flora DDMZ1: application for decolorization of azo dyes

et al. 2022

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Kinetics of Spanish broom peroxidase obeys a Ping-Pong Bi–Bi mechanism with competitive inhibition by substrates

Cited by 28 publications

References 33 publications

Modulation of Aleurone Peroxidases in Kernels of Insect-Resistant Maize (Zea mays L.; Pob84-C3R) After Mechanical and Insect Damage

Modulation of Aleurone Peroxidases in Kernels of Insect-Resistant Maize (Zea mays L.; Pob84-C3R) After Mechanical and Insect Damage

Integrated Experimental and Theoretical Studies on an Electrochemical Immunosensor

Biochemical characterization of a novel azo reductase named BVU5 from the bacterial flora DDMZ1: application for decolorization of azo dyes

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