Sequencing analysis of cDNA clones encoding the American cockroach Cr-PI allergens. Homology with insect hemolymph proteins.
A previous article described the isolation of several lambdagt22A cDNA clones expressing the American cockroach (Periplaneta americana) Cr-PI allergens recognized by both human atopic IgE antibodies and anti-Cr-PI monoclonal antibodies (Wu, C. H., Lee, M. F., and Liao, S. C.(1995) J. Allergy Clin. Immunol. 96, 352-359). This article presents the nucleotide and deduced amino acid sequences of two cDNA clones encoding major allergens of P. americana. Clones C12 and C20 encode proteins of 685 and 631 amino acids with two potential N-glycosylation sites each. The predicted molecular weights for C12 and C20 cloned proteins are 79,300 and 75, 500 with isoelectric point values of 6.26 and 6.63, which are compatible with the determined sizes (Mr 78,000 and 72,000) and isoelectric point value (6.2) of the Cr-PI allergens of P. americana. A high degree of identity (69.1%), including several overlapped predicted central antigenic determinant residues, was found between two allergens. The anti-fusion protein antibody-based enzyme-linked immunosorbent assay was able to detect crude American cockroach extract, Cr-PI, recombinant proteins, and commercial cockroach extracts, which provides further evidence that two allergens share common antigen determinants. Recombinant allergens of clones C12 and C20 both showed 47.4% skin reactivities on 19 cockroach-sensitive asthmatic patients. Unexpectedly, although no sequence similarity was found to other known allergens, two aromatic amino acid-rich allergens were found to have a striking sequence identity to insect storage proteins (20.1-33.9%), insect juvenile hormone-suppressible proteins (30.9-36.4%), and arthropod hemocyanins (29.7-34.6%). Results suggested that two prominent allergens of P. americana are ancestrally related to these insect hemolymph proteins and represent a new group of proteins in the hemocyanin superfamily. These data will now facilitate epitope-mapping studies, and the recombinant allergens may be valuable for diagnostic and therapeutic purposes.
Wu
C H
CH
Department of Medical Research, Taichung Veterans General Hospital, Taiwan 40705.
Lee
M F
MF
Liao
S C
SC
Luo
S F
SF
D000293:Adolescent; D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D001249:Asthma / Q000150:complications; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D000939:Epitopes; D006023:Glycoproteins; D006801:Humans; D006967:Hypersensitivity / Q000150:complications / Q000175:diagnosis; D007301:Insect Hormones / Q000235:genetics; D019476:Insect Proteins; D008875:Middle Aged; D008969:Molecular Sequence Data; D010526:Periplaneta / Q000235:genetics* / Q000276:immunology; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012882:Skin Tests
D018076:DNA, Complementary; D000939:Epitopes; D006023:Glycoproteins; D007301:Insect Hormones; D019476:Insect Proteins; D011506:Proteins; C041137:storage proteins, Insecta; C100630:allergen Cr-PI, Periplaneta americana