cDNA cloning and sequencing reveal the major horse allergen Equ c1 to be a glycoprotein member of the lipocalin superfamily.
The gene encoding the major horse allergen, designated Equus caballus allergen 1 (Equ c1), was cloned from total cDNA of sublingual salivary glands by reverse transcription-polymerase chain reaction using synthetic degenerate oligonucleotides deduced from N-terminal and internal peptide sequences of the glycosylated hair dandruff protein. A recombinant form of the protein, with a polyhistidine tail, was expressed in Escherichia coli and purified by immobilized metal affinity chromatography. The recombinant protein is able to induce a passive cutaneous anaphylaxis reaction in rat, and it behaves similarly to the native Equ c1 in several immunological tests with allergic patients' IgE antibodies, mouse monoclonal antibodies, or rabbit polyclonal IgG antibodies. Amino acid sequence identity of 49-51% with rodent urinary proteins from mice and rats suggests that Equ c1 is a new member of the lipocalin superfamily of hydrophobic ligand-binding proteins that includes several other major allergens. An RNA blot analysis demonstrates the expression of mRNA Equ c1 in liver and in sublingual and submaxillary salivary glands.
Gregoire
C
C
Unité d'Immuno-Allergie, Département de Physiopathologie, Institut Pasteur, 28 rue du Dr Roux, 75024 Paris Cedex 15, France. cgreg@pasteur.fr
Rosinski-Chupin
I
I
Rabillon
J
J
Alzari
P M
PM
David
B
B
Dandeu
J P
JP
D000485:Allergens / Q000235:genetics* / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D006023:Glycoproteins / Q000235:genetics* / Q000276:immunology; D006736:Horses / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D054834:Lipocalins; D008074:Lipoproteins / Q000235:genetics / Q000276:immunology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D005810:Multigene Family; D011994:Recombinant Proteins; D012471:Salivary Proteins and Peptides / Q000235:genetics / Q000276:immunology*; D017386:Sequence Homology, Amino Acid; D012995:Solubility; D014018:Tissue Distribution
D000485:Allergens; D018076:DNA, Complementary; C103025:Equ c1 allergen, Equus caballus; D006023:Glycoproteins; D054834:Lipocalins; D008074:Lipoproteins; D011994:Recombinant Proteins; D012471:Salivary Proteins and Peptides