Molecular cloning of Per a 1 and definition of the cross-reactive Group 1 cockroach allergens

J Allergy Clin Immunol. 1999 May;103(5 Pt 1):859-64. doi: 10.1016/s0091-6749(99)70430-6.

Abstract

Background: Sensitization to allergens produced by German and American cockroaches is strongly associated with the cause of asthma. Most of the cockroach allergens identified to date have been species specific.

Objective: The aim of this study was to identify and sequence cross-reactive cockroach allergens.

Methods: A Periplaneta americana cDNA library was screened with IgE antibody from patients in the United States who were allergic to cockroach and who were sensitized to Blattella germanica.

Results: A cDNA clone was isolated that contained an 870-bp sequence with a 695-bp open reading frame, encoding a 231 amino acid protein, molecular weight 26.2 kd. Plaque immunoassays using anti-Bla g 1 and anti-Per a 1 mAbs and a panel of human IgE antibodies showed that the protein expressed by these clones was Per a 1. Sequence homology searches showed that Per a 1 was homologous to 5 previously reported, but unidentified, sequences from B germanica and P americana. These sequences encoded proteins with multiple molecular sizes containing approximately 100 amino acid repeats. The Per a 1 sequence also showed 31% identity to a mosquito precursor protein, ANG12, which may be involved in digestion. The Per a 1 cDNA was expressed in Pichia pastoris to produce purified recombinant allergen (yield, 14 mg/L).

Conclusion: The results define the molecular structure and antigenic relationships between a new family of cross-reactive "Group 1" allergens produced by both P americana and B germanica. These recombinant allergens and specific mAbs will provide tools to improve the diagnosis and treatment of allergic diseases caused by cockroaches.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Allergens / genetics*
  • Animals
  • Antigens, Plant
  • Base Sequence
  • Cloning, Molecular
  • Cross Reactions / genetics
  • DNA, Complementary / analysis
  • Humans
  • Insect Proteins / genetics
  • Molecular Sequence Data
  • Sequence Homology, Nucleic Acid

Substances

  • ANG12 precursor protein, Anopheles gambiae
  • Allergens
  • Antigens, Plant
  • DNA, Complementary
  • Insect Proteins
  • allergen Per a I