title: [Proteolysis of human proinsulin catalysed by native, modified, and immobilized trypsin].
abstract: Proteolysis of recombinant human proinsulin by the native trypsin, by trypsin modified with a copolymer of vinylpyrrolidone and acrolein, and by the same modified trypsin immobilized on Silochrom 1.5 was studied by RP HPLC and mass spectrometry. Rate constants of the main stages of proinsulin hydrolysis by the native trypsin were estimated. The values of rate constants of the digestions of the most easily hydrolyzable bonds (those formed by the pairs of the basic amino acid residues) in proinsulin were found to be of the same order as those formed by the separate lysine residues (Lys7) and those formed by the four basic amino acid residues of the C-terminal cluster of melittin. It was established that covalent trypsin binding to the copolymer did not change the ratio of the rate constants of the individual stages of proinsulin hydrolysis, whereas after the immobilization of modified trypsin on the Silochrome, the formation of diarginyl insulin-ArgArg, intermediate forms of hydrolyzed insulin, and desThr-insulin proceeds with comparable rates.
authors.forename: O A
authors.initials: OA
authors.lastname: Mirgorodskaia
authors.forename: G A
authors.initials: GA
authors.lastname: Kazanina
authors.forename: E P
authors.initials: EP
authors.lastname: Mirgorodskaia
authors.forename: A A
authors.initials: AA
authors.lastname: Shevchenko
authors.forename: K V
authors.initials: KV
authors.lastname: Mal'tsev
authors.forename: A I
authors.initials: AI
authors.lastname: Miroshnikov
authors.forename: P
authors.initials: P
authors.lastname: Roipstorff
mesh_terms: D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D004800:Enzymes, Immobilized / Q000737:chemistry*; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D008969:Molecular Sequence Data; D011384:Proinsulin / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000737:chemistry*
chemical_list: D004800:Enzymes, Immobilized; D011994:Recombinant Proteins; D011384:Proinsulin; D014357:Trypsin
vernacular_title: Proteoliz proinsulina cheloveka, kataliziruemyĭ nativnym, modifitsirovannym i immobilizirovannym tripsinom.