28MB
Structure of Human Aldehyde oxidase under TCEP-reducing conditions
Summary for 28MB
| Entry DOI | 10.2210/pdb28mb/pdb |
| Descriptor | Aldehyde oxidase, FE2/S2 (INORGANIC) CLUSTER, FLAVIN-ADENINE DINUCLEOTIDE, ... (8 entities in total) |
| Functional Keywords | human aldehyde oxidase, tcep, protein crystallization, oxidoreductase |
| Biological source | Homo sapiens (human) |
| Total number of polymer chains | 2 |
| Total formula weight | 299695.60 |
| Authors | Videira, C.,Esmaeeli, M.,Leimkuhler, S.,Romao, M.J.,Mota, C. (deposition date: 2026-02-06, release date: 2026-07-29, Last modification date: 2026-08-05) |
| Primary citation | Videira, C.,Esmaeeli, M.,Leimkuhler, S.,Romao, M.J.,Mota, C. Structure of human aldehyde oxidase under tris(2-carboxyethyl)phosphine reducing conditions. Acta Crystallogr.,Sect.F, 2026 Cited by PubMed Abstract: The importance of human aldehyde oxidase (hAOX1) has increased in recent decades due to its involvement in drug metabolism. Inhibition studies involving hAOX1 are extensive and a common reducing agent, dithiothreitol (DTT), was recently found to inactivate the enzyme. However, in previous crystallographic studies of hAOX1, DTT was found to be essential for crystallization. To surpass this concern, another reducing agent was used in crystallization trials. Using tris(2-carboxyethyl)phosphine (TCEP), a sulfur-free reducing agent, it was possible to obtain well ordered crystals of wild-type hAOX1 and a variant, hAOX1_6A, which diffracted beyond 2.3 Å resolution. Instead of the typical star-shaped crystals of hAOX1, at pH 4.7 plates are obtained in the orthorhombic space group P222 with two molecules in the asymmetric unit. Activity assays with the enzyme incubated with both reducing agents show that in contrast to DTT, TCEP did not inactivate hAOX1. The replacement of DTT with TCEP in the crystallization of hAOX1 provides a strategy to circumvent enzyme inactivation during crystallographic studies, allowing future applications of new assays, such as time-resolved crystallography. PubMed: 42504845DOI: 10.1107/S2053230X26006904 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.307 Å) |
Structure validation
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