Expression and epitope identification of myosin light chain isoform 1, an allergen in Procambarus clarkii

Food Chem. 2020 Jul 1:317:126422. doi: 10.1016/j.foodchem.2020.126422. Epub 2020 Feb 17.

Abstract

Myosin light chain isoform 1 (MLC1) is reported to be a novel allergen in crayfish (Procambarus clarkii). However, little information is available about its allergic epitopes. In this study, recombinant crayfish MLC1 (rMLC1) was expressed and confirmed by mass spectrometry. Circular dichroic analysis and serological test were performed for the measuring of structural and immunological properties of rMLC1. Specific-protein-A-enriched IgG raised in rabbits against purified rMLC1 was used to screen a phage display random peptide library. Nine MLC1 mimotope clones were identified among 16 random clones after biopanning. Five conformational epitopes were identified with the program LocaPep, and mapped into 3 epitope regions at the antibody-binding interface of MLC1. MLC1 of crayfish showed high primary and secondary structure identity to MLC of other allergenic species, its epitopes were located in the structure conserved regions, and its cross-reactivity among related species was indicated by immunological assays.

Keywords: Cross-reactivity; Epitope; Myosin light chain isoform 1; Phage display; Procambarus clarkii.

MeSH terms

  • Allergens / genetics
  • Allergens / immunology*
  • Allergens / metabolism
  • Amino Acid Sequence
  • Animals
  • Astacoidea / metabolism*
  • Cross Reactions
  • Epitopes / chemistry*
  • Epitopes / immunology
  • Immunoassay
  • Myosin Light Chains / genetics
  • Myosin Light Chains / immunology*
  • Myosin Light Chains / metabolism
  • Peptide Library
  • Protein Isoforms / immunology
  • Protein Isoforms / metabolism
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / immunology
  • Sequence Alignment

Substances

  • Allergens
  • Epitopes
  • Myosin Light Chains
  • Peptide Library
  • Protein Isoforms
  • Recombinant Proteins