Cloning, expression, and primary structure of Metapenaeus ensis tropomyosin, the major heat-stable shrimp allergen

Patrick S C Leung, Ka Hou Chu, Wing Kuen Chow, Aftab Ansari, Claudiu I. Bandea, Hoi Shan Kwan, Stephen M. Nagy, M. Eric Gershwin

Research output: Contribution to journalArticle

204 Scopus citations

Abstract

Shrimp is a common cause of seafood hypersensitivity. To study the mechanism of seafood hypersensitivity at the molecular level, we have determined the primary structure of the major heat-stable allergen of shrimp by cloning, expression, nucleotide sequencing, and amino acid sequence determination of an IgE-reactive cDNA clone, Met e I, isolated from a Metapenaeus ensis expression library in λgt 11. We first constructed a cDNA library from the shrimp M. ensis in λgt 11. We then screened the library with sera from patients with hypersensitivity reactions to shrimp and identified a positive IgE-reactive clone, designated as Met e I. This cDNA was purified to homogeneity and subsequently expressed in the plasmid pGEX. Serum antibodies from patients with shrimp allergy demonstrated positive IgE reactivity by immunoblotting to a protein encoded by the clone Met e I; sera from nonallergic control subjects were not reactive. The nucleotide sequence of this cDNA clone revealed an open reading frame of 281 amino acid residues, coding for a protein of 34 kd. Comparison of the Met e I amino acid sequence with the Genbank database showed that Met e I is highly homologous to multiple isoforms of tropomyosin.

Original languageEnglish (US)
Pages (from-to)882-890
Number of pages9
JournalThe Journal of Allergy and Clinical Immunology
Volume94
Issue number5
DOIs
StatePublished - 1994

Keywords

  • cDNA clone
  • epitope
  • IgE reactivity
  • Metapenaeus ensis
  • recombinant protein
  • Shrimp allergy
  • tropomyosin

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

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