title: Expression of a lipocalin in Pichia pastoris: secretion, purification and binding activity of a recombinant mouse major urinary protein.
abstract: 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.
authors.affiliation: Istituto di Chimica Biologica, Facoltà di Medicina e Chirurgia, Università degli Studi di Parma, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Ferrari
authors.forename: T
authors.initials: T
authors.lastname: Lodi
authors.forename: R T
authors.initials: RT
authors.lastname: Sorbi
authors.forename: R
authors.initials: R
authors.lastname: Tirindelli
authors.forename: A
authors.initials: A
authors.lastname: Cavaggioni
authors.forename: A
authors.initials: A
authors.lastname: Spisni
mesh_terms: D000510:Alpha-Globulins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005822:Genetic Vectors; D051379:Mice; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics*; D011506:Proteins; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D000510:Alpha-Globulins; D018076:DNA, Complementary; D011506:Proteins; D011994:Recombinant Proteins; C035737:major urinary proteins