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Arg354 in the catalytic centre of bovine liver catalase is protected from methylglyoxal-mediated glycation.

Scheckhuber CQ - BMC Res Notes (2015)

Bottom Line: This treatment did not lead to a pronounced reduction of enzymatic activity.Whereas several arginine residues displayed low to moderate levels of glycation (e.g., Arg93, Arg365, Arg444) Arg354 in the active centre of catalase was never found to be modified.Bovine liver catalase is able to tolerate very high levels of the modifying α-oxoaldehyde methylglyoxal so that its essential enzymatic function is not impaired.

View Article: PubMed Central - PubMed

Affiliation: Senckenberg Research Institute, LOEWE Excellence Cluster for Integrative Fungal Research (IPF), Georg-Voigt-Str. 14-16, 60325, Frankfurt, Germany. c.scheckhuber@gmail.com.

ABSTRACT

Background: In addition to controlled post-translational modifications proteins can be modified with highly reactive compounds. Usually this leads to a compromised functionality of the protein. Methylglyoxal is one of the most common agents that attack arginine residues. Methylglyoxal is also regarded as a pro-oxidant that affects cellular redox homeostasis by contributing to the formation of reactive oxygen species. Antioxidant enzymes like catalase are required to protect the cell from oxidative damage. These enzymes are also targets for methylglyoxal-mediated modification which could severely affect their catalytic activity in breaking down reactive oxygen species to less reactive or inert compounds.

Results: Here, bovine liver catalase was incubated with high levels of methylglyoxal to induce its glycation. This treatment did not lead to a pronounced reduction of enzymatic activity. Subsequently methylglyoxal-mediated arginine modifications (hydroimidazolone and dihydroxyimidazolidine) were quantitatively analysed by sensitive nano high performance liquid chromatography/electron spray ionisation/tandem mass spectrometry. Whereas several arginine residues displayed low to moderate levels of glycation (e.g., Arg93, Arg365, Arg444) Arg354 in the active centre of catalase was never found to be modified.

Conclusions: Bovine liver catalase is able to tolerate very high levels of the modifying α-oxoaldehyde methylglyoxal so that its essential enzymatic function is not impaired.

No MeSH data available.


Related in: MedlinePlus

Determination of catalase activity in the absence or presence of methylglyoxal. Mean values of three different measurements are shown in the diagram. Error bars denote the standard deviation of the mean value. MG methylglyoxal (160 mM)
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Fig1: Determination of catalase activity in the absence or presence of methylglyoxal. Mean values of three different measurements are shown in the diagram. Error bars denote the standard deviation of the mean value. MG methylglyoxal (160 mM)

Mentions: A catalase-containing solution that has not been treated with methylglyoxal is capable to degrade half of the hydrogen peroxide present in the assay buffer at time 0 within approx. 45 s (Fig. 1). When the purified catalase enzyme is pre-incubated in buffer containing 160 mM methylglyoxal for 2 h before being diluted into assay buffer no clear differences of enzyme activity compared to the untreated control are measured. Catalase activity is maintained at later time points of the assay (60, 75, 90 s) although it tends to be slightly reduced compared to the untreated control. These data show that bovine liver catalase is able to maintain its activity even in the presence of high concentrations of methylglyoxal.Fig. 1


Arg354 in the catalytic centre of bovine liver catalase is protected from methylglyoxal-mediated glycation.

Scheckhuber CQ - BMC Res Notes (2015)

Determination of catalase activity in the absence or presence of methylglyoxal. Mean values of three different measurements are shown in the diagram. Error bars denote the standard deviation of the mean value. MG methylglyoxal (160 mM)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4696219&req=5

Fig1: Determination of catalase activity in the absence or presence of methylglyoxal. Mean values of three different measurements are shown in the diagram. Error bars denote the standard deviation of the mean value. MG methylglyoxal (160 mM)
Mentions: A catalase-containing solution that has not been treated with methylglyoxal is capable to degrade half of the hydrogen peroxide present in the assay buffer at time 0 within approx. 45 s (Fig. 1). When the purified catalase enzyme is pre-incubated in buffer containing 160 mM methylglyoxal for 2 h before being diluted into assay buffer no clear differences of enzyme activity compared to the untreated control are measured. Catalase activity is maintained at later time points of the assay (60, 75, 90 s) although it tends to be slightly reduced compared to the untreated control. These data show that bovine liver catalase is able to maintain its activity even in the presence of high concentrations of methylglyoxal.Fig. 1

Bottom Line: This treatment did not lead to a pronounced reduction of enzymatic activity.Whereas several arginine residues displayed low to moderate levels of glycation (e.g., Arg93, Arg365, Arg444) Arg354 in the active centre of catalase was never found to be modified.Bovine liver catalase is able to tolerate very high levels of the modifying α-oxoaldehyde methylglyoxal so that its essential enzymatic function is not impaired.

View Article: PubMed Central - PubMed

Affiliation: Senckenberg Research Institute, LOEWE Excellence Cluster for Integrative Fungal Research (IPF), Georg-Voigt-Str. 14-16, 60325, Frankfurt, Germany. c.scheckhuber@gmail.com.

ABSTRACT

Background: In addition to controlled post-translational modifications proteins can be modified with highly reactive compounds. Usually this leads to a compromised functionality of the protein. Methylglyoxal is one of the most common agents that attack arginine residues. Methylglyoxal is also regarded as a pro-oxidant that affects cellular redox homeostasis by contributing to the formation of reactive oxygen species. Antioxidant enzymes like catalase are required to protect the cell from oxidative damage. These enzymes are also targets for methylglyoxal-mediated modification which could severely affect their catalytic activity in breaking down reactive oxygen species to less reactive or inert compounds.

Results: Here, bovine liver catalase was incubated with high levels of methylglyoxal to induce its glycation. This treatment did not lead to a pronounced reduction of enzymatic activity. Subsequently methylglyoxal-mediated arginine modifications (hydroimidazolone and dihydroxyimidazolidine) were quantitatively analysed by sensitive nano high performance liquid chromatography/electron spray ionisation/tandem mass spectrometry. Whereas several arginine residues displayed low to moderate levels of glycation (e.g., Arg93, Arg365, Arg444) Arg354 in the active centre of catalase was never found to be modified.

Conclusions: Bovine liver catalase is able to tolerate very high levels of the modifying α-oxoaldehyde methylglyoxal so that its essential enzymatic function is not impaired.

No MeSH data available.


Related in: MedlinePlus