Int. J. Mol. Sci. 2014, 15(2), 2223-2236; doi:10.3390/ijms15022223
Article

Structural Data on the Periplasmic Aldehyde Oxidoreductase PaoABC from Escherichia coli: SAXS and Preliminary X-ray Crystallography Analysis

1email, 1email, 2email, 3email, 1email, 2email and 1,* email
Received: 25 December 2013; in revised form: 21 January 2014 / Accepted: 22 January 2014 / Published: 31 January 2014
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract: The periplasmic aldehyde oxidoreductase PaoABC from Escherichia coli is a molybdenum enzyme involved in detoxification of aldehydes in the cell. It is an example of an αβγ heterotrimeric enzyme of the xanthine oxidase family of enzymes which does not dimerize via its molybdenum cofactor binding domain. In order to structurally characterize PaoABC, X-ray crystallography and small angle X-ray scattering (SAXS) have been carried out. The protein crystallizes in the presence of 20% (w/v) polyethylene glycol 3350 using the hanging-drop vapour diffusion method. Although crystals were initially twinned, several experiments were done to overcome twinning and lowering the crystallization temperature (293 K to 277 K) was the solution to the problem. The non-twinned crystals used to solve the structure diffract X-rays to beyond 1.80 Å and belong to the C2 space group, with cell parameters a = 109.42 Å, b = 78.08 Å, c = 151.77 Å, β = 99.77°, and one molecule in the asymmetric unit. A molecular replacement solution was found for each subunit separately, using several proteins as search models. SAXS data of PaoABC were also collected showing that, in solution, the protein is also an αβγ heterotrimer.
Keywords: periplasmic aldehyde oxidoreductase; X-ray crystallography; small angle X-ray scattering; crystal twinning
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MDPI and ACS Style

Otrelo-Cardoso, A.R.; da Silva Correia, M.A.; Schwuchow, V.; Svergun, D.I.; Romão, M.J.; Leimkühler, S.; Santos-Silva, T. Structural Data on the Periplasmic Aldehyde Oxidoreductase PaoABC from Escherichia coli: SAXS and Preliminary X-ray Crystallography Analysis. Int. J. Mol. Sci. 2014, 15, 2223-2236.

AMA Style

Otrelo-Cardoso AR, da Silva Correia MA, Schwuchow V, Svergun DI, Romão MJ, Leimkühler S, Santos-Silva T. Structural Data on the Periplasmic Aldehyde Oxidoreductase PaoABC from Escherichia coli: SAXS and Preliminary X-ray Crystallography Analysis. International Journal of Molecular Sciences. 2014; 15(2):2223-2236.

Chicago/Turabian Style

Otrelo-Cardoso, Ana R.; da Silva Correia, Márcia A.; Schwuchow, Viola; Svergun, Dmitri I.; Romão, Maria J.; Leimkühler, Silke; Santos-Silva, Teresa. 2014. "Structural Data on the Periplasmic Aldehyde Oxidoreductase PaoABC from Escherichia coli: SAXS and Preliminary X-ray Crystallography Analysis." Int. J. Mol. Sci. 15, no. 2: 2223-2236.

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