Validation of Recombinant Chicken Liver Bile Acid Binding Protein as a Tool for Cholic Acid Hosting

Biomolecules. 2021 Apr 27;11(5):645. doi: 10.3390/biom11050645.


Bile acids (BAs) are hydroxylated steroids derived from cholesterol that act at the intestinal level to facilitate the absorption of several nutrients and also play a role as signaling molecules. In the liver of various vertebrates, the trafficking of BAs is mediated by bile acid-binding proteins (L-BABPs). The ability to host hydrophobic or amphipathic molecules makes BABPs suitable for the distribution of a variety of physiological and exogenous substances. Thus, BABPs have been proposed as drug carriers, and more recently, they have also been employed to develop innovative nanotechnology and biotechnology systems. Here, we report an efficient protocol for the production, purification, and crystallization of chicken liver BABP (cL-BABP). By means of target expression as His6-tag cL-BABP, we obtained a large amount of pure and homogeneous proteins through a simple purification procedure relying on affinity chromatography. The recombinant cL-BABP showed a raised propensity to crystallize, allowing us to obtain its structure at high resolution and, in turn, assess the structural conservation of the recombinant cL-BABP with respect to the liver-extracted protein. The results support the use of recombinant cL-BABP for the development of drug carriers, nanotechnologies, and innovative synthetic photoswitch systems.

Keywords: X-ray crystallography; bile acids; chicken liver bile acid-binding protein; cholic acid; fatty acid-binding protein; high-resolution crystal structure.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence / genetics
  • Animals
  • Bile Acids and Salts / metabolism
  • Binding Sites / physiology
  • Carrier Proteins / isolation & purification*
  • Carrier Proteins / metabolism
  • Carrier Proteins / pharmacology*
  • Chickens
  • Cholic Acid / analysis
  • Cholic Acid / chemistry
  • Cholic Acid / metabolism
  • Crystallography, X-Ray / methods
  • Drug Delivery Systems / methods*
  • Liver / metabolism
  • Liver / pathology
  • Membrane Glycoproteins / isolation & purification*
  • Membrane Glycoproteins / metabolism
  • Membrane Glycoproteins / pharmacology*
  • Models, Molecular
  • Protein Binding / physiology
  • Recombinant Proteins / metabolism


  • Bile Acids and Salts
  • Carrier Proteins
  • Membrane Glycoproteins
  • Recombinant Proteins
  • bile acid binding proteins
  • Cholic Acid