title: [Identification of the membrane form of phenylalanine hydroxylase from the human liver and characteristics of its subunit composition].
abstract: Phenylalanine hydroxylase was detected among human liver bioptats and autoptats extracted with 0.4% Triton X-100 from the 105,000 g homogenate fraction. In contrast to the membrane form of the rat liver enzyme, human liver phenylalanine hydroxylase is detected both by its enzymatic activity and immunochemically under non-denaturating conditions. The enzymatic activity of phenylalanine hydroxylase makes 5-15% of that of the cytoplasmic fraction and 20-30% of the amount of antigen in the cytoplasmic fraction and 20-30% of the amount of antigen in the cytoplasmic fraction as can be evidenced from rocket immunoelectrophoresis data. Immunoblotting of proteins performed after denaturating electrophoresis of the membrane and cytoplasmic fractions revealed an antigen band with a similar electrophoretic mobility. The subunit composition of the enzyme in both fractions was characterized by two-dimensional electrophoresis with subsequent immunoblotting. It was found that the membrane fraction of the enzyme is represented only by the L-subunit with Mr of 55 kD, whereas the cytoplasmic fraction, besides the predominant L-subunit, also contains 2H-subunits of the enzyme with Mr = 57 kD. These 2H-subunits differ between themselves as well as from the L-subunit by the pI value.
authors: V V
authors: VV
authors: Chestkov
authors: A V
authors: AV
authors: Laptev
authors: S S
authors: SS
authors: Shishkin
mesh_terms: D002462:Cell Membrane / Q000201:enzymology; D003593:Cytoplasm / Q000201:enzymology; D006801:Humans; D008099:Liver / Q000201:enzymology*; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000302:isolation & purification; D010651:Phenylalanine Hydroxylase / Q000302:isolation & purification*; D011487:Protein Conformation; D011489:Protein Denaturation
chemical_list: D046911:Macromolecular Substances; D008565:Membrane Proteins; D010651:Phenylalanine Hydroxylase
vernacular_title: Vyiavlenie membrannoĭ formy fenilalaningidroksilazy pecheni cheloveka i kharakteristika ee sub''edinichnogo sostava.