The bovine plasma retinol-binding protein. Amino acid sequence, interaction with transthyretin, crystallization and preliminary X-ray data.
1. The primary structure of bovine plasma retinol-binding protein (RBP) has been determined and found to be more than 90% identical to human and rabbit RBPs, and more than 80% identical to rat RBP. Main changes in amino acid sequence are observed in two regions on the surface of the protein molecule (residues 138-148 and 169-183). 2. The interactions of bovine RBP with bovine and human transthyretins were investigated using the technique of fluorescence polarization. Bovine RBP was able to form high affinity complexes (K'd = 0.34 +/- 0.02 microM) with both bovine and human transthyretins, displaying a stoichiometry of approximately 2 molecules RBP/molecule transthyretin in both cases. The sites that participate in protein-protein interactions are thus very similar, and this tends to exclude the involvement of the superficial regions more significantly substituted in mammalian RBPs (residues 138-151 and 167-183) in the protein-protein recognition. 3. Bovine RBP has been crystallized (space group P2(1)2(1)2(1), with a = 4.61 nm, b = 4.91 nm, c = 7.61 nm) and the crystals are suitable for high-resolution X-ray diffraction studies.
Institute of Biochemical Sciences, University of Parma, Italy.
R
R
Berni
M
M
Stoppini
M C
MC
Zapponi
M L
ML
Meloni
H L
HL
Monaco
G
G
Zanotti
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D003460:Crystallization; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000302:isolation & purification; D011228:Prealbumin / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D012177:Retinol-Binding Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D054839:Retinol-Binding Proteins, Plasma; D012689:Sequence Homology, Nucleic Acid; D014961:X-Ray Diffraction
D010446:Peptide Fragments; D011228:Prealbumin; D012177:Retinol-Binding Proteins; D054839:Retinol-Binding Proteins, Plasma