Rescue of the orphan enzyme isoguanine deaminase

Daniel S. Hitchcock, Alexander A. Fedorov, Elena V. Fedorov, Lawrence J. Dangott, Steven C. Almo, Frank M. Raushel

Research output: Contribution to journalArticle

7 Citations (Scopus)

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: kcat = 49 s-1, Km = 72 μM, and kcat/Km = 6.7 × 105 M-1 s -1. The kinetic constants for the deamination of cytosine are as follows: kcat = 45 s-1, Km = 302 μM, and kcat/Km = 1.5 × 105 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.

Original languageEnglish (US)
Pages (from-to)5555-5557
Number of pages3
JournalBiochemistry
Volume50
Issue number25
DOIs
StatePublished - Jun 28 2011

Fingerprint

Cytosine Deaminase
Deamination
Enzymes
Escherichia coli
Kinetics
Xanthine
Peptide Mapping
Cytosine
Ammonium Sulfate
Adenine
Fractionation
Chromatography
Gel Chromatography
Anions
isoguanine
Catalytic Domain
Ion exchange
Gels
Oxidation
Peptides

ASJC Scopus subject areas

  • Biochemistry

Cite this

Hitchcock, D. S., Fedorov, A. A., Fedorov, E. V., Dangott, L. J., Almo, S. C., & Raushel, F. M. (2011). Rescue of the orphan enzyme isoguanine deaminase. Biochemistry, 50(25), 5555-5557. https://doi.org/10.1021/bi200680y

Rescue of the orphan enzyme isoguanine deaminase. / Hitchcock, Daniel S.; Fedorov, Alexander A.; Fedorov, Elena V.; Dangott, Lawrence J.; Almo, Steven C.; Raushel, Frank M.

In: Biochemistry, Vol. 50, No. 25, 28.06.2011, p. 5555-5557.

Research output: Contribution to journalArticle

Hitchcock, DS, Fedorov, AA, Fedorov, EV, Dangott, LJ, Almo, SC & Raushel, FM 2011, 'Rescue of the orphan enzyme isoguanine deaminase', Biochemistry, vol. 50, no. 25, pp. 5555-5557. https://doi.org/10.1021/bi200680y
Hitchcock DS, Fedorov AA, Fedorov EV, Dangott LJ, Almo SC, Raushel FM. Rescue of the orphan enzyme isoguanine deaminase. Biochemistry. 2011 Jun 28;50(25):5555-5557. https://doi.org/10.1021/bi200680y
Hitchcock, Daniel S. ; Fedorov, Alexander A. ; Fedorov, Elena V. ; Dangott, Lawrence J. ; Almo, Steven C. ; Raushel, Frank M. / Rescue of the orphan enzyme isoguanine deaminase. In: Biochemistry. 2011 ; Vol. 50, No. 25. pp. 5555-5557.
@article{ec6c6fdbdf0e47eabd539a5a798a498b,
title = "Rescue of the orphan enzyme isoguanine deaminase",
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: kcat = 49 s-1, Km = 72 μM, and kcat/Km = 6.7 × 105 M-1 s -1. The kinetic constants for the deamination of cytosine are as follows: kcat = 45 s-1, Km = 302 μM, and kcat/Km = 1.5 × 105 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.",
author = "Hitchcock, {Daniel S.} and Fedorov, {Alexander A.} and Fedorov, {Elena V.} and Dangott, {Lawrence J.} and Almo, {Steven C.} and Raushel, {Frank M.}",
year = "2011",
month = "6",
day = "28",
doi = "10.1021/bi200680y",
language = "English (US)",
volume = "50",
pages = "5555--5557",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "25",

}

TY - JOUR

T1 - Rescue of the orphan enzyme isoguanine deaminase

AU - Hitchcock, Daniel S.

AU - Fedorov, Alexander A.

AU - Fedorov, Elena V.

AU - Dangott, Lawrence J.

AU - Almo, Steven C.

AU - Raushel, Frank M.

PY - 2011/6/28

Y1 - 2011/6/28

N2 - 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: kcat = 49 s-1, Km = 72 μM, and kcat/Km = 6.7 × 105 M-1 s -1. The kinetic constants for the deamination of cytosine are as follows: kcat = 45 s-1, Km = 302 μM, and kcat/Km = 1.5 × 105 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.

AB - 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: kcat = 49 s-1, Km = 72 μM, and kcat/Km = 6.7 × 105 M-1 s -1. The kinetic constants for the deamination of cytosine are as follows: kcat = 45 s-1, Km = 302 μM, and kcat/Km = 1.5 × 105 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.

UR - http://www.scopus.com/inward/record.url?scp=79959459974&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79959459974&partnerID=8YFLogxK

U2 - 10.1021/bi200680y

DO - 10.1021/bi200680y

M3 - Article

VL - 50

SP - 5555

EP - 5557

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 25

ER -