Structural Analysis of Modified Forms of Recombinant IFN-β Produced under Stress-Simulating Conditions
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Stefania Orrù
Abstract
The present study focused on the investigation of the chemical stability of recombinant human interferon-β (rhIFN-β) 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-β 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-β derivatives showed that rhIFN-β 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.
Copyright © 2000 by Walter de Gruyter GmbH & Co. KG
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Articles in the same Issue
- Enzymatic Halogenation Catalyzed via a Catalytic Triad and by Oxidoreductases
- Structural Analysis of Modified Forms of Recombinant IFN-β Produced under Stress-Simulating Conditions
- A Synthetic Peptide Derived from the Non-Structural Protein 3 of Hepatitis C Virus Serves as a Specific Substrate for PKC
- Neoglycolipids Derived from Phosphatidylethanolamine Serve as Probes in Cell Culture Studies on Glycolipid Metabolism
- Anomalous Binding of MgADP to Myosin of Skeletal Muscle
- Engineering, Purification and Applications of His-Tagged Recombinant Antibody Fragments with Specificity for the Major Birch Pollen Allergen, Bet v1
- Activation of Progelatinase B by Membranes of Human Polymorphonuclear Granulocytes
- Bait Region Cleavage and Complex Formation of Human α2M with a Porphyromonas gingivalisW50 Protease Is Not Accompanied by Enzyme Inhibition
- Adjuvant and Haemolytic Activities of 47 Saponins Derived from Medicinal and Food Plants
- Tissue-Specific Regulation of IRS-2/PI 3-Kinase Association in Aged Rats
- Investigation of the Effect of Freezing on Protease-Catalyzed Peptide Synthesis Using Cryoprotectants and Frozen Organic Solvent
- Cloning and Expression of Functional Equistatin
