Identification and molecular characterization of Charybdis feriatus tropomyosin, the major crab allergen

Patrick S Leung, Yen Chen Chen, M. Eric Gershwin, Shun Hang Wong, Hoi Shan Kwan, Ka Hou Chu

Research output: Contribution to journalArticle

140 Scopus citations

Abstract

Background: Crab sensitivity is one of the most common seafood allergies. However, to date, there has been no report on the molecular characterization of crab allergens and no comparative analysis with other seafood allergens. Objective: This study was undertaken to clone, identify, and determine the primary structure of a major IgE-reactive molecule in crab. Methods: We constructed an expression cDNA library from a common crab, Charybdis feriatus. This library was then screened with the use of sera from subjects with a well-documented history of type I hypersensitivity reactions upon ingestion of crab. An IgE-reactive clone was chosen and subcloned into plasmids for nucleotide sequence determination and expression in Escherichia coli. Results: We identified a 1-kb cDNA designated as Cha f 1. Expression of Cha f 1 produces a 34-kd recombinant protein reactive to the IgE antibodies from patients with crab allergies but not from control subjects. Cha f 1 has an opening reading frame of 264 amino acids and demonstrates marked homology to the shrimp tropomyosin Met e 1. Absorption of allergic sera with Cha f 1 removes IgE reactivity to crab extract. Moreover, absorption of allergic sera with recombinant shrimp Met e 1 tropomyosin removes IgE reactivity to Cha f 1. Conclusions: This 34-kd protein, designated as Cha f 1, is the first identified major allergen of crab. Nucleotide and amino acid comparison shows that this protein is the crab tropomyosin. The molecular basis of shrimp and crab allergy is readily demonstrated at the nucleotide and amino acid level.

Original languageEnglish (US)
Pages (from-to)847-852
Number of pages6
JournalJournal of Allergy and Clinical Immunology
Volume102
Issue number5
StatePublished - 1998

Keywords

  • cDNA
  • Cha f 1
  • Crab allergen
  • Tropomyosin

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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