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.
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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.
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.
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.