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Journal Articles Journal of Biological Chemistry Year : 1994

Pure and functionally homogeneous recombinant retinoid X receptor

Zhi-Ping Chen
  • Function : Author
Lirim Shemshedini
  • Function : Author
Beatrice Durand
  • Function : Author
Noa Noy
  • Function : Author

Abstract

Mouse retinoid X receptor alpha (RXR alpha) lacking the amino-terminal region A/B (RXR alpha delta AB) has been purified to more than 98% purity and functional homogeneity from bacterial and baculovirus-based recombinant expression systems with yields of 2-8 mg/liter of culture. The purified protein is soluble, and fluorescence quenching analysis demonstrated that it binds its cognate ligand 9-cis-retinoic acid (9-cis-RA) stoichiometrically, and with high affinity. Compared with RXR delta AB expressed in COS-1 cells, bacterially and baculovirus-expressed proteins bind approximately 10 and 5 times less efficiently to direct repeat 1 (DR1) DNA elements, respectively, suggesting that animal cell-specific modification of RXR or interaction with other animal cell-specific factors may modulate DNA binding. 9-cis-RA did not stimulate DR1 binding of functional RXR delta AB expressed in Escherichia coli, Sf9 or COS-1 cells. The previously reported ligand effect that can be observed with in vitro made receptor may therefore be a consequence of a conformational stabilization of improperly folded in vitro synthesized protein.

Dates and versions

hal-03831432 , version 1 (26-10-2022)

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Zhi-Ping Chen, Lirim Shemshedini, Beatrice Durand, Noa Noy, Pierre Chambon, et al.. Pure and functionally homogeneous recombinant retinoid X receptor. Journal of Biological Chemistry, 1994, 269 (41), pp.25770-25776. ⟨10.1016/S0021-9258(18)47314-0⟩. ⟨hal-03831432⟩

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