A two-in-one expression construct for biophysical and structural studies of the human pregnane X receptor ligand-binding domain, a pharmaceutical and environmental target.
Carivenc, C., Laconde, G., Blanc, P., Amblard, M., Bourguet, W., Delfosse, V.(2025) Acta Crystallogr F Struct Biol Commun 
- PubMed: 39923198 
- DOI: https://doi.org/10.1107/S2053230X2500069X
- Primary Citation of Related Structures:  
9FZG, 9FZH, 9FZI, 9FZJ - PubMed Abstract: 
The ligand-binding domain (LBD) of the human nuclear receptor pregnane X receptor (PXR) is known to crystallize in two different crystal forms, P2 1 2 1 2 1 or P4 3 2 1 2, depending on the construct and the strategy used for protein production, as well as the presence or absence of the coactivator-derived peptide SRC-1. In order to facilitate biophysical and structural studies, a versatile construct was designed that allows access to both forms. This was achieved by introducing a thrombin cleavage site between the PXR LBD and the SRC-1 peptide fused to its C-terminus. Here, we describe the expression, purification and crystallization processes of this novel construct and report two new structures of PXR LBD that were obtained thanks to this strategy.
Organizational Affiliation: 
Centre de Biologie Structurale (CBS), Univ. Montpellier, INSERM, CNRS, Montpellier, France.