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Chem Biol Interact. 2019 May 1;304:168-172. doi: 10.1016/j.cbi.2019.03.009. Epub 2019 Mar 17.

Expression, purification and crystallization of the novel Xenopus tropicalis ALDH16B1, a homologue of human ALDH16A1.

Author information

1
Department of Pharmacology, School of Medicine, Yale University, New Haven, CT, 06510, USA. Electronic address: georgios.pantouris@yale.edu.
2
Department of Environmental Health Sciences, Yale School of Public Health, New Haven, CT, 06520, USA.
3
Department of Clinical Pharmacy, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado, Aurora, CO, 80045, USA.
4
Department of Environmental Health Sciences, Yale School of Public Health, New Haven, CT, 06520, USA. Electronic address: vasilis.vasiliou@yale.edu.
5
Department of Pharmacology, School of Medicine, Yale University, New Haven, CT, 06510, USA. Electronic address: elias.lolis@yale.edu.

Abstract

ALDH16 is a novel family of the aldehyde dehydrogenase (ALDH) superfamily with unique structural characteristics that distinguish it from the other ALDH superfamily members. In addition to structural characteristics, there is an evolutionary-related grouping within the ALDH 16 genes. The ALDH16 isozymes in frog, lower animals, and bacteria possess a critical Cys residue in their active site, which is absent from ALDH16 in mammals and fish. Genomic analysis and plasma metabolomic studies have associated ALDH16A1 with the pathogenesis of gout in humans, although its actual involvement in this disease is poorly understood. Insight into the structure of ALDH16A1 is an important step in deciphering its function in gout. Herein, we report our efforts towards the structural characterization of Xenopus tropicalis ALDH16B1 (the homolog of human ALDH16A1) that was predicted to be catalytically-active. Recombinant ALDH16B1 was expressed in Sf9 cells and purified using affinity and size exclusion chromatography. Crystallization of ALDH16B1 was achieved by vapor diffusion. A data set was collected at 2.5 Å and preliminary crystallographic analysis showed that the frog ALDH16B1 crystals belong to the P 212 121 space group with unit cell parameters a = 80.48 Å, b = 89.73 Å, c = 190.92 Å, α = β = γ = 90.00°. Structure determination is currently in progress.

KEYWORDS:

Aldehyde dehydrogenase; Catalytic residue; Crystallization; Frog ALDH16B1; Human ALDH16A1

PMID:
30894314
DOI:
10.1016/j.cbi.2019.03.009
[Indexed for MEDLINE]

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