Purification and characterization of parvalbumins, the major allergens in red stingray (Dasyatis akajei)

J Agric Food Chem. 2010 Dec 22;58(24):12964-9. doi: 10.1021/jf103316h. Epub 2010 Dec 1.


Fish has received increasing attention because it induces IgE-mediated food allergy. Parvalbumin (PV) represents the major allergen of fish, and IgE cross-reactivity to PV in various teleost fish species has been shown, while little information is available about allergens in elasmobranch fish. In this study, two PV isoforms (named as PV-I and PV-II) from red stingray (Dasyatis akajei) were purified to homogeneity by a series of procedures including ammonium sulfate precipitation and column chromatographies of DEAE-Sepharose and Sephacryl S-200. Purified PVs revealed a single band on tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The molecular masses of PV-I and PV-II were 12.29 and 11.95 kDa, respectively, as determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Western blot using antifrog PV monoclonal antibody (PARV-19) showed positive reactions to the two proteins, confirming that they were PVs, although their immunological reactivities were weaker than those of PV from silver carp. The N-terminal amino acid sequence of PV-I was determined, and comparison with PVs from other fish species showed low homology between teleost and elasmobranch fish. The isoelectric points of PV-I and PV-II were 5.4 and 5.0, respectively, as determined by two-dimensional electrophoresis (2-DE), suggesting that both isoforms belong to the α-group. IgE immunoblotting analysis showed that sera from fish-allergic patients reacted to both PV-I and PV-II from red stingray. Thermal stability revealed that PV-I easily formed oligomers than PV-II, which might contribute to the maintenance of its allerginicity during heat processing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allergens / chemistry*
  • Allergens / immunology
  • Allergens / isolation & purification*
  • Amino Acid Sequence
  • Animals
  • Fish Proteins / chemistry*
  • Fish Proteins / immunology
  • Fish Proteins / isolation & purification*
  • Molecular Sequence Data
  • Molecular Weight
  • Parvalbumins / chemistry*
  • Parvalbumins / immunology
  • Parvalbumins / isolation & purification*
  • Protein Stability
  • Sequence Alignment
  • Skates, Fish / immunology*


  • Allergens
  • Fish Proteins
  • Parvalbumins