Three-Dimensional Structure of the Immunophilin-like Domain of FKBP59 in Solution

• Published in: Biochemistry (Easton) 1996-08, Vol.35 (34), p.11045-11052
• Main Authors: Craescu, Constantin T, Rouvière, Nathalie, Popescu, Aurel, Cerpolini, Esther, Lebeau, Marie-Claire, Baulieu, Etienne-Emile, Mispelter, Joël
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Carrier Proteins - chemistry; Carrier Proteins - metabolism; Cattle; Crystallography, X-Ray; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - metabolism; Heat-Shock Proteins - chemistry; Heat-Shock Proteins - metabolism; Humans; Hydrogen Bonding; Immunosuppressive Agents - metabolism; Immunosuppressive Agents - pharmacology; Magnetic Resonance Spectroscopy; Models, Molecular; Molecular Sequence Data; Polyenes - metabolism; Polyenes - pharmacology; Protein Conformation; Protein Structure, Secondary; Protein Structure, Tertiary; Sequence Homology, Amino Acid; Sirolimus; Tacrolimus - metabolism; Tacrolimus - pharmacology; Tacrolimus Binding Proteins
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi960975p

FKBP59 is a protein usually associated with heat-shock protein hsp90 and steroid receptors. The N-terminal domain of the rabbit liver protein (149 amino acids) has a sequence homology with FKBP12, binds FK506 immunosuppressor, and has a peptidyl−prolyl cis−trans isomerase activity. The three-dimensional structure of this domain (FKBP59-I) was determined using homo- and heteronuclear multidimensional NMR spectroscopy, distance geometry, and molecular dynamics methods. Structure calculations used 1290 interproton distance restraints derived from nuclear Overhauser enhancement measurements, 29 dihedral φ angle restraints, and 92 hydrogen bond restraints. For the final 22 structures, the root mean square distance from the mean atomic coordinates, calculated for well-defined secondary structure fragments, is 0.47 ± 0.05 and 1.26 ± 0.15 Å for backbone heavy atoms (N, Cα, C‘) and for all non-hydrogen atoms, respectively. The global fold contains a twisted six-stranded antiparallel β-sheet and a short α-helix packed on the hydrophobic side of the sheet. The 20 N-terminal and 12 C-terminal amino acids of the domain are disordered. The main-chain structure of FKBP59-I is globally similar to the NMR-derived and X-ray structures of unbound FKBP12. An unusual hydrogen bond interaction between the indole amino proton of Trp 89 and the aromatic cycle of Phe 129 was observed. This gives a large upfield shift (−4.8 ppm) and a significant exchange protection factor. The implications of the present structure determination on the ligand binding of FKBP59 are discussed.