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Liver-type fatty acid binding protein interacts with hepatocyte nuclear factor 4α
Avery L McIntosh1, Anca D Petrescu, Heather A Hostetler
1Department of Physiology and Pharmacology, Texas A&M University, TVMC, College Station, TX 77843-4466, United States.
Liver type fatty acid binding protein (L-FABP) interacts with Hepatocyte nuclear factor 4α (HNF4α), influencing its activity. This study reveals L-FABP binds HNF4α, altering its structure and enhancing its function in the nucleus.
Area of Science:
- Molecular Biology
- Biochemistry
- Cell Biology
Background:
- Hepatocyte nuclear factor 4α (HNF4α) is a key regulator of gene expression in the liver.
- Liver-type fatty acid binding protein (L-FABP) is known to be regulated by HNF4α.
Purpose of the Study:
- To investigate the reciprocal interaction between L-FABP and HNF4α.
- To elucidate the functional consequences of this interaction on HNF4α activity.
Main Methods:
- Fluorescence resonance energy transfer (FRET) microscopy to assess proximity and binding affinity (Kd).
- Circular dichroism (CD) spectroscopy to analyze changes in protein secondary structure.
- Reporter gene assays in COS7 cells to evaluate HNF4α transactivation.
Main Results:
- L-FABP and HNF4α were found in close proximity (~80 Å) with high binding affinity (Kd ~250-300 nM).
- The interaction between L-FABP and HNF4α resulted in alterations to protein secondary structure.
- L-FABP potentiated the transactivation activity of HNF4α in cellular assays.
Conclusions:
- L-FABP directly interacts with HNF4α, both structurally and functionally.
- This interaction modulates HNF4α's secondary structure and enhances its transcriptional activity.
- L-FABP acts as a signaling molecule, contributing to HNF4α activation within the nucleus.
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