Abstract
In the present work the effect of a mutation on tyrosine 33 residue (Y33G) of human cytidine deaminase (CDA) was investigated with regard to protein solubility and specific activity. Osmolytes and CDA ligands were used to increase the yield and the specific activity of the protein. The mutant enzyme was purified and subjected to a kinetic characterization and to stability studies. These investigations reinforced the hypothesis that in human CDA the side chain of Y33 is involved in intersubunit interactions with four glutamate residues (E108) forming a double latch that connects each of the two pairs of monomers of the tetrameric CDA.
Copyright © 2010 Elsevier B.V. All rights reserved.
Publication types
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Research Support, N.I.H., Intramural
MeSH terms
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Animals
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Blotting, Western
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Catalytic Domain
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Circular Dichroism
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Cytidine Deaminase / antagonists & inhibitors
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Cytidine Deaminase / chemistry*
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Cytidine Deaminase / isolation & purification
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Cytidine Deaminase / metabolism*
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Electrophoresis, Polyacrylamide Gel
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Enzyme Stability / drug effects
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Humans
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Kinetics
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Mice
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Molecular Chaperones / pharmacology
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Mutant Proteins / antagonists & inhibitors
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Mutant Proteins / isolation & purification
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Mutant Proteins / metabolism
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Protein Structure, Quaternary
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Protein Structure, Secondary
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Protein Unfolding / drug effects
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Structure-Activity Relationship
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Temperature
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Tyrosine / metabolism*
Substances
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Molecular Chaperones
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Mutant Proteins
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Tyrosine
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Cytidine Deaminase