title: Structural analysis of modified forms of recombinant IFN-beta produced under stress-simulating conditions.
abstract: The present study focused on the investigation of the chemical stability of recombinant human interferon-beta (rhIFN-beta) tested in vitro by chemical treatments that simulate stress-induced conditions that may occur during handling, storage or ageing of protein samples. Mild oxidation and/or alkylation of the recombinant protein showed that the four methionines occurring in the interferon displayed different chemical susceptibility in that Met36 and Met117 were fully modified, whereas Met1 showed only little modification and Met62 was completely resistant. Moreover, incubation of rhIFN-beta under alkaline conditions resulted in the formation of a covalent dimeric species stabilised by an intermolecular disulphide bridge involving the free SH group of Cys17 from each polypeptide chain. Analysis of biological activity of the different IFN-beta derivatives showed that rhIFN-beta fully retains its specific activity following mild oxidation treatments whereas reaction with a high concentration of alkylating agents or incubation under alkaline conditions strongly reduce its specific antiviral activity.
authors.affiliation: Centro Internazionale Servizi di Spettrometria di Massa, CNR-Università di Napoli, Federico II, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Orrù
authors.forename: A
authors.initials: A
authors.lastname: Amoresano
authors.forename: R
authors.initials: R
authors.lastname: Siciliano
authors.forename: R
authors.initials: R
authors.lastname: Napoleoni
authors.forename: O
authors.initials: O
authors.lastname: Finocchiaro
authors.forename: A
authors.initials: A
authors.lastname: Datola
authors.forename: E
authors.initials: E
authors.lastname: De Luca
authors.forename: A
authors.initials: A
authors.lastname: Sirna
authors.forename: P
authors.initials: P
authors.lastname: Pucci
mesh_terms: D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D016899:Interferon-beta / Q000096:biosynthesis / Q000737:chemistry* / Q000494:pharmacology; D013058:Mass Spectrometry; D018384:Oxidative Stress; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology
chemical_list: D000998:Antiviral Agents; D011994:Recombinant Proteins; D016899:Interferon-beta