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Ligand binding properties of human cellular retinoic acid binding protein II expressed in E. coli as a glutathione‐ S ‐transferase fusion protein
Author(s) -
Redfern Christopher P.F.,
Wilson Katherine E.
Publication year - 1993
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(93)80100-9
Subject(s) - retinoic acid , fusion protein , ligand (biochemistry) , biochemistry , glutathione , retinoic acid receptor , chemistry , glutathione s transferase , transferase , microbiology and biotechnology , biology , enzyme , recombinant dna , receptor , gene
To test the hypothesis that 9‐ cis ‐retinoic acid is a ligand for cellular retinoic acid binding protein II (CRABPII), human CRABPII was expressed as a glutathione‐ S ‐transferase fusion protein (GST‐CRABP II) and a single affinity purification step used to extract it from bacterial lysates. GST‐CRABP II bound all trans ‐retinoic acid with high affinity ( K d 14.2 ± 6.5 nM), but 9‐ cis ‐retinoic acid bound poorly. These studies suggest that 9‐ cis ‐retinoic acid is not a ligand for CRABP II. Their ease of purification makes GST‐CRABP fusion proteins useful tools for ligand binding studies with different retinoids.