Cargando…

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

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 doma...

Descripción completa

Detalles Bibliográficos
Autores principales: Otrelo-Cardoso, Ana Rita, da Silva Correia, Márcia Alexandra, Schwuchow, Viola, Svergun, Dmitri I., Romão, Maria João, Leimkühler, Silke, Santos-Silva, Teresa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Diversity Preservation International (MDPI) 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3958847/
https://www.ncbi.nlm.nih.gov/pubmed/24492481
http://dx.doi.org/10.3390/ijms15022223
_version_ 1782307955015680000
author Otrelo-Cardoso, Ana Rita
da Silva Correia, Márcia Alexandra
Schwuchow, Viola
Svergun, Dmitri I.
Romão, Maria João
Leimkühler, Silke
Santos-Silva, Teresa
author_facet Otrelo-Cardoso, Ana Rita
da Silva Correia, Márcia Alexandra
Schwuchow, Viola
Svergun, Dmitri I.
Romão, Maria João
Leimkühler, Silke
Santos-Silva, Teresa
author_sort Otrelo-Cardoso, Ana Rita
collection PubMed
description 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.
format Online
Article
Text
id pubmed-3958847
institution National Center for Biotechnology Information
language English
publishDate 2014
publisher Molecular Diversity Preservation International (MDPI)
record_format MEDLINE/PubMed
spelling pubmed-39588472014-03-20 Structural Data on the Periplasmic Aldehyde Oxidoreductase PaoABC from Escherichia coli: SAXS and Preliminary X-ray Crystallography Analysis Otrelo-Cardoso, Ana Rita da Silva Correia, Márcia Alexandra Schwuchow, Viola Svergun, Dmitri I. Romão, Maria João Leimkühler, Silke Santos-Silva, Teresa Int J Mol Sci Article 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. Molecular Diversity Preservation International (MDPI) 2014-01-31 /pmc/articles/PMC3958847/ /pubmed/24492481 http://dx.doi.org/10.3390/ijms15022223 Text en © 2014 by the authors; licensee MDPI, Basel, Switzerland http://creativecommons.org/licenses/by/3.0/ This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Otrelo-Cardoso, Ana Rita
da Silva Correia, Márcia Alexandra
Schwuchow, Viola
Svergun, Dmitri I.
Romão, Maria João
Leimkühler, Silke
Santos-Silva, Teresa
Structural Data on the Periplasmic Aldehyde Oxidoreductase PaoABC from Escherichia coli: SAXS and Preliminary X-ray Crystallography Analysis
title Structural Data on the Periplasmic Aldehyde Oxidoreductase PaoABC from Escherichia coli: SAXS and Preliminary X-ray Crystallography Analysis
title_full Structural Data on the Periplasmic Aldehyde Oxidoreductase PaoABC from Escherichia coli: SAXS and Preliminary X-ray Crystallography Analysis
title_fullStr Structural Data on the Periplasmic Aldehyde Oxidoreductase PaoABC from Escherichia coli: SAXS and Preliminary X-ray Crystallography Analysis
title_full_unstemmed Structural Data on the Periplasmic Aldehyde Oxidoreductase PaoABC from Escherichia coli: SAXS and Preliminary X-ray Crystallography Analysis
title_short Structural Data on the Periplasmic Aldehyde Oxidoreductase PaoABC from Escherichia coli: SAXS and Preliminary X-ray Crystallography Analysis
title_sort structural data on the periplasmic aldehyde oxidoreductase paoabc from escherichia coli: saxs and preliminary x-ray crystallography analysis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3958847/
https://www.ncbi.nlm.nih.gov/pubmed/24492481
http://dx.doi.org/10.3390/ijms15022223
work_keys_str_mv AT otrelocardosoanarita structuraldataontheperiplasmicaldehydeoxidoreductasepaoabcfromescherichiacolisaxsandpreliminaryxraycrystallographyanalysis
AT dasilvacorreiamarciaalexandra structuraldataontheperiplasmicaldehydeoxidoreductasepaoabcfromescherichiacolisaxsandpreliminaryxraycrystallographyanalysis
AT schwuchowviola structuraldataontheperiplasmicaldehydeoxidoreductasepaoabcfromescherichiacolisaxsandpreliminaryxraycrystallographyanalysis
AT svergundmitrii structuraldataontheperiplasmicaldehydeoxidoreductasepaoabcfromescherichiacolisaxsandpreliminaryxraycrystallographyanalysis
AT romaomariajoao structuraldataontheperiplasmicaldehydeoxidoreductasepaoabcfromescherichiacolisaxsandpreliminaryxraycrystallographyanalysis
AT leimkuhlersilke structuraldataontheperiplasmicaldehydeoxidoreductasepaoabcfromescherichiacolisaxsandpreliminaryxraycrystallographyanalysis
AT santossilvateresa structuraldataontheperiplasmicaldehydeoxidoreductasepaoabcfromescherichiacolisaxsandpreliminaryxraycrystallographyanalysis