Peroxidase improves the activity of catalase by preventing aggregation during TFE-induced denaturation.
Catalase, a ubiquitous enzyme of the free radical scavenging machinery unfolds and aggregates in the presence of 2,2,2-triflouroethanol (TFE). Catalase molecule aggregates at 50% TFE as evident by high thioflavin T fluorescence, shifted congo red absorbance, change in circular dichroism and soret spectra. TEM images confirmed the nature of catalase aggregates to be oligomers. Organic solvent-induced aggregation of catalase is prevented by the presence of peroxidase (another enzyme of the free radical scavenging machinery). To alter the progress of aggregation in presence of increasing concentration of TFE, we determined the effect of peroxidase on catalase oligomerization by several different techniques, including turbidity measurement, activity assay, thioflavin T fluorescence, circular dichroism, shift in congo red absorbance, transmission electron microscopy (TEM), Rayleigh scattering, soret absorption spectra, and ANS fluorescence. The presence of peroxidase in the vicinity of folded catalase helps it to remain functionally active and inhibited aggregation in the presence of TFE, suggesting that proteins are stable in crowded environments. Moreover, this catalase-peroxidase interaction is biologically significant as it provides insights into how the aggregation process may be altered.
a Faculty of Life Sciences, Department of Biochemistry , Aligarh Muslim University , Aligarh 202002 , India.
Mohammad
M
Furkan
a Faculty of Life Sciences, Department of Biochemistry , Aligarh Muslim University , Aligarh 202002 , India.
Asim
A
Rizvi
a Faculty of Life Sciences, Department of Biochemistry , Aligarh Muslim University , Aligarh 202002 , India.
Md Tauqir
MT
Alam
a Faculty of Life Sciences, Department of Biochemistry , Aligarh Muslim University , Aligarh 202002 , India.
Aabgeena
A
Naeem
D002374:Catalase / Q000737:chemistry*; D002942:Circular Dichroism; D003224:Congo Red; D005023:Ethylene Chlorohydrin / Q000031:analogs & derivatives / Q000737:chemistry; D016166:Free Radical Scavengers / Q000737:chemistry; D009195:Peroxidase / Q000737:chemistry*; D066329:Protein Aggregates / Q000187:drug effects*; D011485:Protein Binding; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation; D017510:Protein Folding; D055503:Protein Multimerization / Q000187:drug effects*; D013050:Spectrometry, Fluorescence
D016166:Free Radical Scavengers; D066329:Protein Aggregates; D003224:Congo Red; D005023:Ethylene Chlorohydrin; C005849:2,2,2-trichloroethanol; D002374:Catalase; D009195:Peroxidase
TFE; aggregation; catalse; peroxidase; techniques