NR AJVW
AU Ramoni,R.; Vincent,F.; Ashcroft,A.E.; Accornero,P.; Grolli,S.; Valencia,C.; Tegoni,M.; Cambillau,C.
TI Control of domain swapping in bovine odorant-binding protein
QU Biochemical Journal 2002 Aug 1; 365(3): 739-48
PT journal article
AB As revealed by the X-ray structure, bovine odorant-binding protein (OBPb) is a domain swapped dimer [Tegoni, Ramoni, Bignetti, Spinelli and Cambillau (1996) Nat. Struct. Biol. 3, 863-867; Bianchet, Bains, Petosi, Pevsner, Snyder, Monaco and Amzel (1996) Nat. Struct. Biol. 3, 934-939]. This contrasts with all known mammalian OBPs, which are monomers, and in particular with porcine OBP (OBPp), sharing 42.3% identity with OBPb. By the mechanism of domain swapping, monomers are proposed to evolve into dimers and oligomers, as observed in human prion. Comparison of bovine and porcine OBP sequences pointed at OBPp glycine 121, in the hinge linking the beta-barrel to the alpha-helix. The absence of this residue in OBPb might explain why the normal lipocalin beta-turn is not formed. In order to decipher the domain swapping determinants we have produced a mutant of OBPb in which a glycine residue was inserted after position 121, and a mutant of OBPp in which glycine 121 was deleted. The latter mutation did not result in dimerization, while OBPb-121Gly+ became monomeric, suggesting that domain swapping was reversed. Careful structural analysis revealed that besides the presence of a glycine in the hinge, the dimer interface formed by the C-termini and by the presence of the lipocalins conserved disulphide bridge may also control domain swapping.
MH Amino Acid Sequence; Animal; Anthracenes/metabolism; Cattle; Crystallography, X-Ray; Dimerization; Fluorescent Dyes/metabolism; Human; Models, Molecular; Molecular Sequence Data; Molecular Weight; Mutagenesis, Site-Directed; Protein Binding; Protein Isoforms; *Protein Structure, Tertiary; Receptors, Odorant/*chemistry/genetics/metabolism; Recombinant Proteins/genetics/metabolism; Sequence Alignment; Support, Non-U.S. Gov't; Swine
AD Dipartimento di Produzioni Animali, Biotecnologie Veterinaire, Qualita e Sicurezza degli Alimenti, Universita di Parma, Via del Taglio 8, 43100 Parma, Italy.
SP englisch
PO England