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Metal binding to cutinase-like enzyme from Saccharomonospora viridis AHK190 and its effects on enzyme activity and stability
- Source :
- The Journal of Biochemistry.
- Publication Year :
- 2019
- Publisher :
- Oxford University Press (OUP), 2019.
-
Abstract
- A cutinase from Saccharomonospora viridis AHK190, Cut190, can hydrolyze polyethylene terephthalate and has a unique feature that the activity and stability are regulated by Ca2+ binding. Our recent structural and functional analyses showed three Ca2+ binding sites and their respective roles. Here, we analysed the binding thermodynamics of Mn2+, Zn2+ and Mg2+ to Cut190 and their effects on the catalytic activity and thermal stability. The binding affinities of Mn2+ and Zn2+ were higher than that of Mg2+ and are all entropy driven with a binding stoichiometry of three, one and one for Zn2+, Mn2+ and Mg2+, respectively. The catalytic activity was measured in the presence of the respective metals, where the activity of 0.25 mM Mn2+ was comparable to that of 2.5 mM Ca2+. Our 3D Reference Interaction Site Model calculations suggested that all the ions exhibited a high occupancy rate for Site 2. Thus, Mn2+ and Mg2+ would most likely bind to Site 2 (contributes to stability) with high affinity, while to Sites 1 and 3 (contributes to activity) with low affinity. We elucidate the metal-dependent structural and functional properties of Cut190 and show the subtle balance on structure stability and flexibility is controlled by specific metal ions.
- Subjects :
- Models, Molecular
inorganic chemicals
Cutinase
Circular dichroism
Stereochemistry
Metal ions in aqueous solution
Biochemistry
03 medical and health sciences
Enzyme Stability
Magnesium
Thermal stability
Binding site
Molecular Biology
030304 developmental biology
chemistry.chemical_classification
Manganese
0303 health sciences
Binding Sites
biology
Chemistry
030302 biochemistry & molecular biology
Isothermal titration calorimetry
General Medicine
Enzyme assay
Actinobacteria
Zinc
Enzyme
Mutation
biology.protein
Thermodynamics
Calcium
Carboxylic Ester Hydrolases
Subjects
Details
- ISSN :
- 17562651 and 0021924X
- Database :
- OpenAIRE
- Journal :
- The Journal of Biochemistry
- Accession number :
- edsair.doi.dedup.....148923a62acaece783750ece8c49da45