Solution structure of a recombinant mouse major urinary protein

Major urinary proteins (MUPs) form an ensemble of protein isoforms which ar e expressed and secreted by sexually mature male mice only. They belong to the lipocalin superfamily and share with other members of this family the c apacity to bind hydrophobic molecules, some of which are odorants. MUPs, ei ther associated with or free of their natural ligands, play an important ro le in the reproductive cycle of these rodents by acting as pheromones. In f act, they are able to interact with receptors in the vomeronasal organ of t he female mice, inducing hormonal and physiological responses by an as yet unknown mechanism. In order to investigate the structural and dynamical features of these prot eins in solution, one of the various wild-type isoforms (rMUP: 162 residues ) was cloned and subsequently isotopically labeled. The complete H-1, C-13 and N-15 resonance assignment of that isoform, achieved by using a variety of multidimensional heteronuclear NMR experiments, has been reported recent ly. Here, we describe the refined high-resolution three-dimensional solution st ructure of rMUP in the native state, obtained by a combination of distance geometry and energy minimization calculations based on 2362 NOE-derived dis tance restraints. A comparison with the crystal structure of the wild-type MUPs reveals, aside from minor differences, a close resemblance in both sec ondary structure and overall topology. The secondary structure of the protein consists of eight antiparallel beta- strands forming a single beta-sheet and an alpha-helix in the C-terminal re gion. In addition, there are several helical and hairpin turns distributed throughout the protein sequence, mostly connecting the beta-strands. The te rtiary fold of the beta-sheet creates a beta-barrel, common to all members of the lipocalin superfamily. The shape of the beta-barrel resembles a caly x, lined inside by mostly hydrophobic residues that are instrumental for th e binding and transport of small nonpolar ligand molecules.