2HLV

Bovine Odorant Binding Protein deswapped triple mutant

Summary for 2HLV

• Entry DOI: 10.2210/pdb2hlv/pdb
• Related: 1OBP
• Descriptor: Odorant-binding protein, 3,6-BIS(METHYLENE)DECANOIC ACID, GLYCEROL, ... (4 entities in total)
• Functional Keywords: odorant binding protein, domain swapping, transport protein
• Biological source: Bos taurus (cattle)
• Cellular location: Secreted: P07435
• Total number of polymer chains: 1
• Total formula weight: 18756.73

Authors

Spinelli, S.,Tegoni, M.,Cambillau, C. (deposition date: 2006-07-10, release date: 2008-03-25, Last modification date: 2024-10-09)

Primary citation

Ramoni, R.,Spinelli, S.,Grolli, S.,Conti, V.,Merli, E.,Cambillau, C.,Tegoni, M.
Deswapping bovine odorant binding protein.
Biochim.Biophys.Acta, 1784:651-657, 2008

PubMed Abstract: The X-ray structure of bovine Odorant Binding Protein (bOBP) revealed its association as a domain swapped dimer. bOBP, devoid of any cysteines, contrasts with other mammalian OBPs, which are monomeric and possess at least one disulfide bridge. We have produced a mutant of bOBP in which a glycine residue was inserted after position 121. This mutation yielded a monomeric bOBP-121Gly+ in which domain swapping has been reverted. Here, we have subsequently introduced two mutations, Trp64Cys and His155Cys, in view to stabilize the putative monomer with a disulfide bridge. We have determined the crystal structure of this triple mutant at 1.65 A resolution. The mutant protein is monomeric, stabilized by a disulfide bridge between Trp64Cys and His155Cys, with a backbone superimposable to that of native bOBP, with the exception of the hinge and of the 10 residues at the C-terminus. bOBP triple mutant binds 1-amino-anthracene, 1-octen-3-ol (bOBP co-purified ligand) and other ligands with microM Kd values comparable to those of the swapped dimer.

PubMed: 18269920
DOI: 10.1016/j.bbapap.2008.01.010

Experimental method

X-RAY DIFFRACTION (1.65 Å)