Poster Session I

Project Type

Poster

Project Funding and Affiliations

University of Montana Grant 25-0339 “Characterizing PPARg Interactions with Intrinsically Disordered Coregulator Domains Using Analytical Ultracentrifugation” National Institutes of Health Grant 1R01DK129646 Center for Biomolecular Structure and Dynamics, NIH Grant S10-OD036366

Faculty Mentor’s Full Name

Borries Demeler

Faculty Mentor’s Department

Analytical Ultracentrifugation

Abstract / Artist's Statement

Coregulator proteins bind to nuclear receptors, thereby modulating transcription activity. Coregulators are required by these nuclear receptors for efficient transcriptional regulation. [2] The equilibrium dissociation constant (KD) of coregulator proteins to nuclear receptors can be measured in solution using techniques such as isothermal calorimetry (ITC) and fluorescence anisotropy (FA). Coregulators' ability to self associate is relevant in how nuclear receptors are activated. Furthermore, self-association of coregulators comprises the modeling reliability of ITC and FA data, thus introducing errors into ITC and FA data. [3].

The Hughes Lab has purified various coregulator proteins to be used for in vitro experimentation.

Using sedimentation velocity analytical ultracentrifugation, we aim to characterize coregulator self-association and obtain relative KD values. This information will validate other ligand modulated coregulator binding experimentation, and aid in better understanding how these ligand-regulated nuclear receptors become activated within the cell. The ability for these proteins to oligomerize will guide future studies regarding the phase-separation of coregulators and their respective transcription complexes. In this poster, we use coregulator CBP 1-127 as an example of our experimental process.

Category

Life Sciences

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Apr 17th, 10:45 AM Apr 17th, 11:45 AM

Coactivator Self-Oligomerization Through AUC

UC South Ballroom

Coregulator proteins bind to nuclear receptors, thereby modulating transcription activity. Coregulators are required by these nuclear receptors for efficient transcriptional regulation. [2] The equilibrium dissociation constant (KD) of coregulator proteins to nuclear receptors can be measured in solution using techniques such as isothermal calorimetry (ITC) and fluorescence anisotropy (FA). Coregulators' ability to self associate is relevant in how nuclear receptors are activated. Furthermore, self-association of coregulators comprises the modeling reliability of ITC and FA data, thus introducing errors into ITC and FA data. [3].

The Hughes Lab has purified various coregulator proteins to be used for in vitro experimentation.

Using sedimentation velocity analytical ultracentrifugation, we aim to characterize coregulator self-association and obtain relative KD values. This information will validate other ligand modulated coregulator binding experimentation, and aid in better understanding how these ligand-regulated nuclear receptors become activated within the cell. The ability for these proteins to oligomerize will guide future studies regarding the phase-separation of coregulators and their respective transcription complexes. In this poster, we use coregulator CBP 1-127 as an example of our experimental process.