Molecular and allergenic properties of natural hemocyanin from Chinese mitten crab (Eriocheir sinensis)

Hemocyanin in crustaceans is an allergen for humans. However, little information was available on its molecular, structural and allergenic properties. In this study, the purified natural protein was identified as Eriocheir sinensis HC by LC-MS/MS, which was allergenic because its reaction with the serum IgE of crustacean patients. Results of the molecular properties showed that, HC was resistant to trypsin digestion, but not a heat-stable protein. Boiling (55.05 ± 3.50 %) and steaming (66.84 ± 1.65 %) induced an increase in β-sheet and decreased allergenicity of HC. By comparing the amino acid sequences of eight crustaceans, HC was found to be highly conserved. Five epitopes of HC were identified and validated by murine sensitization model, and two of them (P3 and P10) were exactly as the predicted by six types of bioinformatics. Multiple bioinformatics analysis combining with murine sensitization model seemed to be effective way for identification of allergenic epitopes.