title: Insertion of a casein kinase recognition sequence induces phosphorylation of ovine beta-lactoglobulin in transgenic mice.
abstract: We have shown that the cellular mechanisms of the mammary gland can be used to produce a phosphorylated form of a normally unphosphorylated milk protein. This was achieved by the insertion of a beta-casein DNA sequence coding for a group of mammary gland casein kinase recognition sites into ovine beta-lactoglobulin. Transgenic mice carrying this modified gene were generated and lactating females were shown to produce a novel beta-lactoglobulin in their milk. The infrared spectrum, reactivity to antiphosphoserine antibody and reduction of electrophoretic mobility on treatment with alkaline phosphatase showed that the novel protein recovered from the milk whey (serum) was phosphorylated and molecular mass determination by mass spectrometry was consistent with the phosphorylation of one or two residues. A similar level of phosphorylation was measured by quantitative infrared spectroscopy. Centrifugation of the milk to pellet the casein micelles showed that most of the phosphorylated beta-lactoglobulin was in the whey and hence not incorporated into casein micelles.
authors.affiliation: Roslin Institute Edinburgh, Midlothian, UK.
authors.forename: M
authors.initials: M
authors.lastname: McClenaghan
authors.forename: E
authors.initials: E
authors.lastname: Hitchin
authors.forename: E M
authors.initials: EM
authors.lastname: Stevenson
authors.forename: A J
authors.initials: AJ
authors.lastname: Clark
authors.forename: C
authors.initials: C
authors.lastname: Holt
authors.forename: J
authors.initials: J
authors.lastname: Leaver
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D047388:Casein Kinases; D017931:DNA Primers; D007782:Lactoglobulins / Q000378:metabolism*; D008321:Mammary Glands, Animal / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D010766:Phosphorylation; D011494:Protein Kinases / Q000737:chemistry / Q000378:metabolism*; D012756:Sheep
chemical_list: D017931:DNA Primers; D007782:Lactoglobulins; D011494:Protein Kinases; D047388:Casein Kinases