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Cloning and sequencing of the Dermatophagoides pteronyssinus group III allergen, Der p III
Author(s) -
SMITH W.A.,
CHUA K.Y.,
KUO M.C.,
ROGERS B. L.,
THOMAS W. R.
Publication year - 1994
Publication title -
clinical and experimental allergy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.462
H-Index - 154
eISSN - 1365-2222
pISSN - 0954-7894
DOI - 10.1111/j.1365-2222.1994.tb00223.x
Subject(s) - complementary dna , allergen , house dust mite , cdna library , microbiology and biotechnology , peptide sequence , gel electrophoresis , amino acid , biology , recombinant dna , chemistry , biochemistry , allergy , immunology , gene
Summary House dust mites are widely recognized as major factors involved in the triggering of allergic diseases such as asthma. It is now apparent that the group III allergens of the Dermatophagoides mite species may play a significant role in a number of house dust mite allergic cases. Natural Der p III was isolated by gel filtration of salt precipitated Dermatophagoides pteronyssinus extract and as reported previously ran as a doublet of Mr 28 and 30 K on sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE). Natural Der f III was isolated by affinity purification with the 5A12 monoclonal antibody. Amino acid sequence data was generated for both these proteins which was used to construct DNA probes to screen a Dermatophagoides pteronyssinus cDNA library by hybridization and resulted in the isolation of a recombinant Der p III cDNA clone, P3WSI. The 1059 bp cDNA fragment included a 786 bp open reading frame which encodes a pre‐pro region of 29 amino acids and a mature protein of 232 amino acids with a calculated Mr 24985. A search of the BLAST protein database has confirmed that the Der p III P3WS1 clone is approximately 50% homologous with other trypsin proteins. We have confirmed with both our natural protein sequence and the P3WS1 amino acid sequence data that the group III allergens are trypsin‐like proteins.