Major Urinary Protein Complex Research Articles

Glycosylated major urinary protein of the house mouse: characterization of its N-linked oligosaccharides.

A minor component of the major urinary protein complex of the house mouse was chromatographically isolated and ascertained to be a previously suspected glycoprotein. Using highly sensitive mass-spectrometric techniques for sequencing and linkage analysis, the N-linked oligosaccharides of this glycoprotein were characterized. They were determined to be of the complex type with a wide heterogeneity. The heterogeneity was due to both the degree of sialylation and the presence of galactose residues in either beta(1-3) or beta(1-4) linkages. The biantennary structures were the most pronounced glycans, while tri- and tetraantennary entities were minor.

Expression of a lipocalin in Pichia pastoris: secretion, purification and binding activity of a recombinant mouse major urinary protein

The proteins of the mouse major urinary protein complex (MUP), members of the lipocalin family, bind volatile pheromones and interact with the vomeronasal neuroepithelium of the olfactory system. We report the expression of a MUP protein using its native signal sequence for secretion in the methylotrophic yeast, Pichia pastoris. Mature recombinant MUP (rMUP) is secreted at a concentration of 270 mg/l in minimal medium and it is isolated from the culture supernatant by one step ion-exchange chromatography in a nearly pure form. Binding activity, tested with an odorant molecule which displays high affinity for native MUP, indicates that rMUP has a behavior similar to the native one. This finding suggests that the protein, and in particular its hydrophobic binding pocket, is properly folded.

Acceleration of puberty onset in female mice by male urinary proteins.

1. Puberty onset in female mice is accelerated by exposure to conspecific adult male urine, which acts through the vomeronasal organ and the accessory olfactory system. A distinctive component of adult male mouse urine is the major urinary protein complex (MUP), which is a lipocalin; it has a hydrophobic pocket that binds small endogenous volatile molecules. The MUP gene family also codes for a hexapeptide, which has four residues in common with the N-terminal region of MUP. 2. MUP, the volatiles bound to MUP and the MUP-related hexapeptide have been tested for the induction of puberty acceleration by measuring the increase in uterus weight related to the first pro-oestrus phase. MUP, together with its bound volatiles, induces puberty acceleration. Its activity is retained even when the volatiles have been removed either by organic extraction or competition displacement with a high-affinity ligand. 3. MUP-related hexapeptide also induces puberty acceleration in female mice. In contrast, the odorants bound to MUP do not exert this effect. 4. It is proposed that the vomeronasal organ contains receptors that recognize the short N-terminal consensus sequence, N-Glu-Glu-Ala-X-Ser (where X is a polar residue), common to both MUP and the hexapeptide.

Characterization of allergens from urine and pelts of laboratory mice

Antigens from CBA/H, BALB/c and C57BL mouse urine and CBA/H mouse pelts were studied to identify allergens that sensitize laboratory workers exposed to mice. Potent allergens in mouse urine were identified within the major urinary protein (MUP)‡ complex, which was found to have a molecular weight of 16,500 daltons (unreduced) and 22,000 daltons (reduced). The isoelectric point of the major component was 4.9. CBA/H, BALB/c and C57BL urine also contained material of low molecular weight (< 15,000 daltons) which was only slightly less potent than MUP allergens, but was present in relatively small amounts. In the three mouse strains, purified MUP proteins that bound to IgE antibodies cross-reacted extensively with one another, and with allergens in dust from a mouse room. There was complete antigenic cross-reactivity between urinary protein of pI 4.9 and low molecular weight fractions of CBA/H pelt antigens at the same isoelectnc point, suggesting that part of the allergenicity of pelt material may result from its content of components of the MUP complex. However, other allergens with a higher molecular weight (62,000 daltons) and isoelectric points between 5.5 and 6.7 were identified by gel filtration chromatography and preparative isoelectric focusing of CBA/H pelt material.

Regulation by androgen of mRNA level for the major urinary protein complex in mouse liver

Summary RNA extracted from mouse liver directed the synthesis of the major urinary protein (MUP) complex in a heterologous cell-free system derived from wheat germ. Livers of normal males and dihydrotestosterone-treated females showed much higher level of mRNA for the MUP complex than those of normal females, indicating that androgen controls the synthesis of the MUP complex at pretranslational level in liver.

Simplified procedure for electrophoresis of the major urinary protein of Mus musculus.

Simplified procedure for electrophoresis of the major urinary protein of Mus musculus.

Components of the major urinary protein complex in inbred mice: separation and peptide mapping.

Gel filtration of the nondialyzable fraction of urine from normal inbred mice on Sephadex G-100 yielded three peaks (I, II, and III in order of elution), the relative sizes of which varied with the sex and strain of the mice. Constituents of peak I, the breakthrough peak, included uromucoid (Tamm-Horsfall mucoprotein); peak III was low in nitrogen, rich in carbohydrate, nonprecipitable with trichloroacetic acid, gave no definitive ultraviolet or visible spectrum, and had a sedimentation coefficient of 0.5 S. Peak II contained the electrophoretically distinguishable prealbumins of the major urinary protein (MUP) complex. These components (known as 1, 2, and 3 in order of increasing mobility toward the anode) were separated by chromatography on diethylaminoethyl cellulose. Tryptic peptide mapping indicated that components 1 and 2, a genetic variant shown to be under the control of one genetic locus (the Mup-alocus), differed by a single peptide. Components 1 and 3 had a number of peptides in common plus several peptides unique to each. The peptide map of any given component did not differ between sexes or between the strains investigated.

Linkage of one component of the major urinary protein complex of mice to the brown coat color locus

Two codominant alleles,Mup-a1andMup-a2, controlling electrophoretic variation of one of the components of the major urinary protein (MUP) complex of the mouseMus musculushave been found to be linked to the black–brown coat color alleles (linkage group VIII).

Major urinary protein complex of normal mice: origin.

Mouse serum contains protein having the same charge density and molecular size as the major urinary protein complex of mice. Mouse liver (but not eight other tissues examined) incorporated amino acids labeled with carbon-14 into the complex in vitro. The degree of incorporation was greater in livers from males than from females, and was internmediate in livers from females treated with testosterone.

Electrophoretic Variation and Sex Dimorphism of the Major Urinary Protein Complex in Inbred Mice: A New Genetic Marker

Electrophoretic Variation and Sex Dimorphism of the Major Urinary Protein Complex in Inbred Mice: A New Genetic Marker