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Purify cytidine deaminase from Escherichia coli B and evaluate its catalytic properties, pH stability, substrate range, and inhibitor interactions.

December 1, 1971Journal of Biological ChemistryOpen Access

Cytidine Deaminase from Escherichia coli

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RCRobert M. CohenCincinnati VA Medical Center RWRichard WolfendenUniversity of North Carolina at Chapel Hill

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Implication

Biochemical analysis reveals potent inhibition of cytidine deaminase by tetrahydrouridine in Escherichia coli, highlighting an unusually tight enzyme-inhibitor binding affinity.

Key Points

Purify cytidine deaminase from Escherichia coli B and evaluate its catalytic properties, pH stability, substrate range, and inhibitor interactions.
Purified cytidine deaminase approximately 160-fold from crude extracts of Escherichia coli B.
Measured catalytic activity across pH 6 to 11, evaluated solvent isotope effects using D2O, and tested alternative substrate hydrolysis with N4-methylcytidine.
Assessed competitive inhibition using 3,4,5,6-tetrahydrouridine and compared inhibitor affinity to reaction product affinity.
The enzyme retains constant catalytic activity between pH 6 and 11, displaying no significant rate alteration when D2O replaces water as the solvent.
Purified deaminase mediates the deamination of cytidine and also catalyzes the slow hydrolysis of N4-methylcytidine.
3,4,5,6-Tetrahydrouridine competitively inhibits the enzyme with an affinity exceeding the combined affinity for products uridine and ammonia by more than four orders of magnitude.

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Cohen et al. (1971) studied this question.

synapsesocial.com/papers/6a70c6e18031ec7bb1dc90e6 https://doi.org/10.1016/s0021-9258(19)45812-2

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Implication

Biochemical analysis reveals potent inhibition of cytidine deaminase by tetrahydrouridine in Escherichia coli, highlighting an unusually tight enzyme-inhibitor binding affinity.

Cytidine Deaminase from Escherichia coli

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Key Points

Cite This Study

Cytidine Deaminase from Escherichia coli

Discussion

Member takes

Implication

Key Points

Cite This Study

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