Tropomyosin, a myofibrillar muscle protein, has been recognized as a finfish allergen. In this study, tropomyosin from Atlantic cod fillets (Gadus morhua, CTM) was purified using a two-step purification strategy (isoelectric precipitation and anion-exchange chromatography). CTM structural configuration in two sample matrices (impure and pure) were elaborated using different polyacrylamide gel electrophoresis (native, non-reducing, and reducing PAGE). Their corresponding immunoblots were conducted to investigate CTM antigenicity under three conditions. Overall, CTM retained solubility, integrity, and antigenicity after heat treatment. Three CTM monomeric α-type isoforms (33 kDa) were identified using two-dimensional PAGE. Under native condition, the vast majority of CTM existed in the disulfide-reduced dimeric form (66 kDa). Under non-reducing condition, sodium dodecyl sulfate (anionic surfactant) broke CTM dimers, leaving monomers and disulfide-induced tetramers. Under reducing condition, β-mercaptoethanol (thiol reducing agent) dissociated disulfide-linked CTM tetramers (132 kDa) into monomers (33 kDa). CTM retained antigenicity regardless of structural configuration under different conditions.
Keywords: Disulfide-induced tetramer; Disulfide-reduced dimer; Finfish; Structural configuration; Tropomyosin; α-type isoforms.
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