Identification of a Novel Cat Allergen – Cystatin

Abstract

Background: Cat allergen is an important cause of sensitization among children with asthma in Japan. Although there is good evidence that cats produce other allergens, only one major allergen, Fel d 1, has been studied in detail. Aims: To identify and define the molecular structure of the other potential cat allergens. Methods: A cat skin cDNA library was screened using IgE antibodies to cat dander and selected clones were sequenced and expressed. Results: One cDNA clone contained an open reading frame encoding a 98-amino acid residue protein. Sequence homology searches revealed a high degree of identity with bovine and human cystatin A, 79 and 75%, respectively. This cat cystatin clone contained the conserved cysteine protease motif and two of three lipocalin motifs. By plaque immunoassay, 60–90% of cat allergic sera had IgE Ab to cat cystatin. This cysteine protease inhibitor motif was partially conserved in dog allergens, Can f 1 and Can f 2, which are lipocalins. Recombinant cystatin was produced in Escherichia coli cells and purified as an 11-kD protein, corresponding to the predicted MW of cystatin. The structure of cat cystatin was modeled on human cystatin B using the SWISS-MODEL. Conclusion: A newly identified allergen, cystatin, has been cloned from cat skin and is a member of the cysteine protease inhibitor family.