Probing the conformational dynamics of full length PR interactions with TBP using HDX

摘要

1. HDX analysis of full length PR constructs reveal the highly dynamic nature (intrinsic disorder) of the N-terminal domain. 2. Agonist (R5020) bound full length PR-B showed strongest interaction with TBPc. Several regions in the LBD (including AF2) showed stabilization while interacting with TBPc. On the other hand, R5020 bound full length PR-A showed protection only in the AF2 region. PR-B is believed to be the more transcriptionally active isoform which is consistent with these data. 3. Effect of functional class of ligand, agonist and antagonist, on PR-TBP interaction was observed in by HDX analysis of full length PR. 4. TBPc has stronger interaction with full length PR-B compare to full length PR-A suggesting that PR-B interacts more strongly with TBP than does PR-A. 5. Both isolated N-terminal domains of PR-A and PR-B (NTD-A and NTD-B) confer stabilization at the same region of TBPc. NTD-B induces destabilization at the region of AA 119-137 in TBPc. 6. Agonist bound truncated PR-B (243-933) construct showed an intermediate (between strong and weak) level of interaction with TBPc.