title: The role of interchain disulfide bond in a recombinant human interleukin-17A variant.
abstract: Interleukin-17A (IL-17A) is the prototype of IL-17 family and has been implicated in the pathogenesis of a variety of autoimmune diseases. Therefore its structural and functional properties are of great medical interest. During our research on a recombinant human IL-17A (rhIL-17A) variant, four isoforms were obtained when it was refolded. While isoforms 1 and 2 represented non-covalent dimers, isoforms 3 and 4 were determined to be covalent dimers. All four isoforms were structurally similar by Circular Dichroism and fluorescence spectroscopy studies, but differential scanning calorimetry demonstrated thermal stability in the order of isoform 1=isoform 2<isoform 4<isoform 3. In addition, compared to covalent dimers (isoform 3 and 4), the non-covalent dimers (isoforms 1 and 2) are slightly less active in a receptor-binding assay but at least 5-fold less active in a cell-based assay.
authors.affiliation: Biologics Research, Janssen Research and Development, LLC., 1400 McKean Road, Spring House, PA 19477, United States. Electronic address: bwu63@its.jnj.com.
authors.forename: Bingyuan
authors.initials: B
authors.lastname: Wu
authors.affiliation: Biologics Research, Janssen Research and Development, LLC., 1400 McKean Road, Spring House, PA 19477, United States.
authors.forename: Salman
authors.initials: S
authors.lastname: Muzammil
authors.affiliation: Immunology Discovery Research, Janssen Research and Development, LLC., 1400 McKean Road, Spring House, PA 19477, United States.
authors.forename: Brian
authors.initials: B
authors.lastname: Jones
authors.affiliation: Biologics Research, Janssen Research and Development, LLC., 1400 McKean Road, Spring House, PA 19477, United States.
authors.forename: Jennifer F
authors.initials: JF
authors.lastname: Nemeth
authors.affiliation: Biologics Research, Janssen Research and Development, LLC., 1400 McKean Road, Spring House, PA 19477, United States.
authors.forename: Dariusz J
authors.initials: DJ
authors.lastname: Janecki
authors.affiliation: Biologics Research, Janssen Research and Development, LLC., 1400 McKean Road, Spring House, PA 19477, United States.
authors.forename: Audrey
authors.initials: A
authors.lastname: Baker
authors.affiliation: Immunology Discovery Research, Janssen Research and Development, LLC., 1400 McKean Road, Spring House, PA 19477, United States.
authors.forename: M
authors.initials: M
authors.lastname: Merle Elloso
authors.affiliation: Biologics Research, Janssen Research and Development, LLC., 1400 McKean Road, Spring House, PA 19477, United States.
authors.forename: Michael
authors.initials: M
authors.lastname: Naso
authors.affiliation: Biologics Research, Janssen Research and Development, LLC., 1400 McKean Road, Spring House, PA 19477, United States.
authors.forename: Jill
authors.initials: J
authors.lastname: Carton
authors.affiliation: Biologics Research, Janssen Research and Development, LLC., 1400 McKean Road, Spring House, PA 19477, United States.
authors.forename: Susann
authors.initials: S
authors.lastname: Taudte
mesh_terms: D000595:Amino Acid Sequence; D002152:Calorimetry, Differential Scanning; D002942:Circular Dichroism; D004220:Disulfides / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D020381:Interleukin-17 / Q000737:chemistry / Q000378:metabolism*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D020033:Protein Isoforms / Q000378:metabolism; D055503:Protein Multimerization; D058849:Protein Refolding; D011994:Recombinant Proteins / Q000378:metabolism*; D012996:Solutions; D013050:Spectrometry, Fluorescence
chemical_list: D004220:Disulfides; C508717:IL17A protein, human; D020381:Interleukin-17; D010455:Peptides; D020033:Protein Isoforms; D011994:Recombinant Proteins; D012996:Solutions
keywords: Activity; CD; CHCA; Circular Dichroism; DSC; Disulfide bond; HPBA; IL-17A; MMmeas; MMtheo; PrOH; Refolding; alpha-cyano-4-hydroxycinnamic acid; differential scanning calorimetry; heptafluoro-butyric anhydride; measured molar mass; propanol; theoretical molar mass