3RN6

Crystal structure of Cytosine Deaminase from Escherichia Coli complexed with zinc and isoguanine

Summary for 3RN6

• Entry DOI: 10.2210/pdb3rn6/pdb
• Related: 3O7U
• Descriptor: Cytosine deaminase, (2S)-1-[3-{[(2R)-2-hydroxypropyl]oxy}-2,2-bis({[(2R)-2-hydroxypropyl]oxy}methyl)propoxy]propan-2-ol, ZINC ION, ... (5 entities in total)
• Functional Keywords: amidohydrolase fold, cytosine deaminase, isoguanine, hydrolase
• Biological source: Escherichia coli
• Total number of polymer chains: 1
• Total formula weight: 48295.19

Authors

Fedorov, A.A.,Fedorov, E.V.,Hitchcock, D.S.,Raushel, F.M.,Almo, S.C. (deposition date: 2011-04-22, release date: 2011-08-24, Last modification date: 2023-09-13)

Primary citation

Hitchcock, D.S.,Fedorov, A.A.,Fedorov, E.V.,Dangott, L.J.,Almo, S.C.,Raushel, F.M.
Rescue of the orphan enzyme isoguanine deaminase.
Biochemistry, 50:5555-5557, 2011

PubMed Abstract: Cytosine deaminase (CDA) from Escherichia coli was shown to catalyze the deamination of isoguanine (2-oxoadenine) to xanthine. Isoguanine is an oxidation product of adenine in DNA that is mutagenic to the cell. The isoguanine deaminase activity in E. coli was partially purified by ammonium sulfate fractionation, gel filtration, and anion exchange chromatography. The active protein was identified by peptide mass fingerprint analysis as cytosine deaminase. The kinetic constants for the deamination of isoguanine at pH 7.7 are as follows: k(cat) = 49 s(-1), K(m) = 72 μM, and k(cat)/K(m) = 6.7 × 10(5) M(-1) s(-1). The kinetic constants for the deamination of cytosine are as follows: k(cat) = 45 s(-1), K(m) = 302 μM, and k(cat)/K(m) = 1.5 × 10(5) M(-1) s(-1). Under these reaction conditions, isoguanine is the better substrate for cytosine deaminase. The three-dimensional structure of CDA was determined with isoguanine in the active site.

PubMed: 21604715
DOI: 10.1021/bi200680y

Experimental method

X-RAY DIFFRACTION (2.255 Å)