Analysis of amino acid sequence variations and immunoglobulin E-binding epitopes of German cockroach tropomyosin

Kyoung Yong Jeong, Jongweon Lee, In Yong Lee, Han Il Ree, Chein Soo Hong, Tai Soon Yong

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The allergenicities of tropomyosins from different organisms have been reported to vary. The cDNA encoding German cockroach tropomyosin (Bla g 7) was isolated, expressed, and characterized previously. In the present study, the amino acid sequence variations in German cockroach tropomyosin were analyzed in order to investigate its influence on allergenicity. We also undertook the identification of immunodominant peptides containing immunoglobulin E (IgE) epitopes which may facilitate the development of diagnostic and immunotherapeutic strategies based on the recombinant proteins. Two-dimensional gel electrophoresis and immunoblot analysis with mouse anti-recombinant German cockroach tropomyosin serum was performed to investigate the isoforms at the protein level. Reverse transcriptase PCR (RT-PCR) was applied to examine the sequence diversity. Eleven different variants of the deduced amino acid sequences were identified by RT-PCR. German cockroach tropomyosin has only minor sequence variations that did not seem to affect its allergenicity significantly. These results support the molecular basis underlying the cross-reactivities of arthropod tropomyosins. Recombinant fragments were also generated by PCR, and IgE-binding epitopes were assessed by enzyme-linked immunosorbent assay. Sera from seven patients revealed heterogeneous IgE-binding responses. This study demonstrates multiple IgE-binding epitope regions in a single molecule, suggesting that full-length tropomyosin should be used for the development of diagnostic and therapeutic reagents.

Original languageEnglish
Pages (from-to)874-878
Number of pages5
JournalClinical and Diagnostic Laboratory Immunology
Volume11
Issue number5
DOIs
Publication statusPublished - 2004 Sep 1

Fingerprint

Blattellidae
Tropomyosin
Protein Sequence Analysis
Immunoglobulin E
Epitopes
Amino Acids
RNA-Directed DNA Polymerase
Reverse Transcriptase Polymerase Chain Reaction
Amino Acid Sequence
Immunosorbents
Arthropods
Electrophoresis, Gel, Two-Dimensional
Electrophoresis
Serum
Recombinant Proteins
Assays
Protein Isoforms
Complementary DNA
Gels
Enzyme-Linked Immunosorbent Assay

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology
  • Clinical Biochemistry
  • Microbiology (medical)

Cite this

Jeong, Kyoung Yong ; Lee, Jongweon ; Lee, In Yong ; Ree, Han Il ; Hong, Chein Soo ; Yong, Tai Soon. / Analysis of amino acid sequence variations and immunoglobulin E-binding epitopes of German cockroach tropomyosin. In: Clinical and Diagnostic Laboratory Immunology. 2004 ; Vol. 11, No. 5. pp. 874-878.
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Analysis of amino acid sequence variations and immunoglobulin E-binding epitopes of German cockroach tropomyosin. / Jeong, Kyoung Yong; Lee, Jongweon; Lee, In Yong; Ree, Han Il; Hong, Chein Soo; Yong, Tai Soon.

In: Clinical and Diagnostic Laboratory Immunology, Vol. 11, No. 5, 01.09.2004, p. 874-878.

Research output: Contribution to journalArticle

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