The Refined Structure of dUTPase from Escherichia coli
(1998) In Acta Crystallographica. Section D: Biological Crystallography D54(5). p.735-749- Abstract
- Deoxyuridine 5'-triphosphate nucleotidohydrolase (dUTPase, E.C. 3.6.1.23) catalyzes the hydrolysis of dUTP to dUMP and pyrophosphate and is involved in nucleotide metabolism and DNA synthesis. A crystal of the recombinant E. coli enzyme, precipitated from polyethylene glycol mixtures in the presence of succinate at pH 4.2, was used to collect synchrotron diffraction data to 1.9 Å resolution, in space group R3, a = b = 86.62, c = 62.23 Å. Mercury and platinum derivative data were collected at wavelengths to optimize the anomalous contribution. The resulting 2.2 Å MIRAS phases differed from the final set by 40° on average and produced an excellent map which was easy to interpret. The model contains 132 water molecules and refined to an R... (More)
- Deoxyuridine 5'-triphosphate nucleotidohydrolase (dUTPase, E.C. 3.6.1.23) catalyzes the hydrolysis of dUTP to dUMP and pyrophosphate and is involved in nucleotide metabolism and DNA synthesis. A crystal of the recombinant E. coli enzyme, precipitated from polyethylene glycol mixtures in the presence of succinate at pH 4.2, was used to collect synchrotron diffraction data to 1.9 Å resolution, in space group R3, a = b = 86.62, c = 62.23 Å. Mercury and platinum derivative data were collected at wavelengths to optimize the anomalous contribution. The resulting 2.2 Å MIRAS phases differed from the final set by 40° on average and produced an excellent map which was easy to interpret. The model contains 132 water molecules and refined to an R value of 13.7%. 136 residues have clear electron density out of 152 expected from the gene sequence. The 16 C-terminal residues are presumably disordered in the crystal lattice. The monomer is a `jelly-roll' type, containing mostly -sheet and only one short helix. The molecule is a tight trimer. A long C-terminal arm extends from one subunit and encompasses the next one within the trimer contributing to its -sheet. Conserved sequence motifs common among dUTPases, previously suggested to compose the active site and confirmed in a recent study of the dUDP complex, are located at subunit-subunit interfaces along the threefold axis, in parts of the -sheet and in loop regions. A similar molecular architecture has recently been found in two other trimeric dUTPases. (Less)
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https://lup.lub.lu.se/record/125499
- author
- Dauter, Z ; Wilson, K S ; Larsson, G ; Nyman, Per-Olof LU and Cedergren-Zeppezauer, E S
- organization
- publishing date
- 1998
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Acta Crystallographica. Section D: Biological Crystallography
- volume
- D54
- issue
- 5
- pages
- 735 - 749
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- scopus:0032167573
- ISSN
- 1399-0047
- DOI
- 10.1107/S0907444997016223
- language
- English
- LU publication?
- yes
- id
- 9b304292-f41f-4b6c-98eb-ed7a915007f4 (old id 125499)
- date added to LUP
- 2016-04-01 16:13:49
- date last changed
- 2022-01-28 18:12:16
@article{9b304292-f41f-4b6c-98eb-ed7a915007f4, abstract = {{Deoxyuridine 5'-triphosphate nucleotidohydrolase (dUTPase, E.C. 3.6.1.23) catalyzes the hydrolysis of dUTP to dUMP and pyrophosphate and is involved in nucleotide metabolism and DNA synthesis. A crystal of the recombinant E. coli enzyme, precipitated from polyethylene glycol mixtures in the presence of succinate at pH 4.2, was used to collect synchrotron diffraction data to 1.9 Å resolution, in space group R3, a = b = 86.62, c = 62.23 Å. Mercury and platinum derivative data were collected at wavelengths to optimize the anomalous contribution. The resulting 2.2 Å MIRAS phases differed from the final set by 40° on average and produced an excellent map which was easy to interpret. The model contains 132 water molecules and refined to an R value of 13.7%. 136 residues have clear electron density out of 152 expected from the gene sequence. The 16 C-terminal residues are presumably disordered in the crystal lattice. The monomer is a `jelly-roll' type, containing mostly -sheet and only one short helix. The molecule is a tight trimer. A long C-terminal arm extends from one subunit and encompasses the next one within the trimer contributing to its -sheet. Conserved sequence motifs common among dUTPases, previously suggested to compose the active site and confirmed in a recent study of the dUDP complex, are located at subunit-subunit interfaces along the threefold axis, in parts of the -sheet and in loop regions. A similar molecular architecture has recently been found in two other trimeric dUTPases.}}, author = {{Dauter, Z and Wilson, K S and Larsson, G and Nyman, Per-Olof and Cedergren-Zeppezauer, E S}}, issn = {{1399-0047}}, language = {{eng}}, number = {{5}}, pages = {{735--749}}, publisher = {{John Wiley & Sons Inc.}}, series = {{Acta Crystallographica. Section D: Biological Crystallography}}, title = {{The Refined Structure of dUTPase from Escherichia coli}}, url = {{http://dx.doi.org/10.1107/S0907444997016223}}, doi = {{10.1107/S0907444997016223}}, volume = {{D54}}, year = {{1998}}, }