title: Characterization of lipovitellin 2 as a tyrosine-phosphorylated protein in oocytes, eggs and early embryos of Xenopus laevis.
abstract: A tyrosine-phosphorylated protein of 33 kDa is shown to be present in the solubilized yolk fraction of Xenopus laevis oocytes, eggs, and early embryos. The phosphoprotein is lipovitellin 2 as demonstrated by immunoprecipitation and mass spectrometry studies, and is termed pp33/LV2. Sub-cellular fractionation and immunoblotting studies demonstrate that pp33/LV2 is stably present in the Triton X-100-resistant and SDS-soluble yolk fractions during oogenesis, oocyte maturation, and early embryogenesis. From after the swimming tadpole stages (stage 39∼), however, it becomes partly soluble to Triton X-100-containing buffer and all disappear thereafter (stage 48∼49). In vitro enzyme assays with the use of the tyrosine phosphatase LAR and the tyrosine kinase Src demonstrate the reversible nature of the tyrosine phosphorylation of pp33/LV2. Microinjection studies demonstrate that the solubilized yolk fractions, but not those immunodepleted of pp33/LV2 or those pretreated with LAR, inhibit progesterone- or insulin-induced oocyte maturation. A pp33/LV2-like protein seems to present in two Xenopus subspecies, one other frog species, and two fish species, but not in other amphibian species, such as newt and salamander. These results suggest that LV2, in its tyrosine-phosphorylated form, serves in a cellular function in a species-specific manner, but the mechanism is still unknown.
authors.affiliation: The Graduate School of Engineering, Kyoto Sangyo University, Kyoto, Japan.
authors.forename: Shota
authors.initials: S
authors.lastname: Kushima
authors.forename: Gunay
authors.initials: G
authors.lastname: Mammadova
authors.forename: A K M
authors.initials: AK
authors.lastname: Mahbub Hasan
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Fukami
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Sato
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015387:Egg Proteins, Dietary / Q000378:metabolism*; D004625:Embryo, Nonmammalian / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism*; D010766:Phosphorylation; D014981:Xenopus / Q000196:embryology* / Q000378:metabolism
chemical_list: D015387:Egg Proteins, Dietary; C106338:lipovitellin 2