title: Molecular cloning and the allergenic characterization of tropomyosin from Tyrophagus putrescentiae.
abstract: Storage mites have been recognized as a cause of asthma and rhinitis. Studies from several countries have shown that the IgE-mediated allergy to storage mites is of considerable importance, especially in rural populations. This study aimed to identify and characterize new allergens from Tyrophagus putrescentiae. A partial cDNA sequence encoding tropomyosin was isolated from the cDNA library by immunoscreening using anti-mouse IgG1 sera raised against T. putrescentiae whole body extract. The deduced amino acid sequence shares 64-94% identity with previously known allergenic tropomyosins. Its recombinant protein was produced by using a pET 28b expression system and purified by affinity chromatography using Ni-NTA agarose. The IgE reactivities of tropomyosins from T. putrescentiae and Dermatophagoides farinae were compared by enzyme linked immunosorbent assay (ELISA). Recombinant Tyr p 10 showed 12.5% (5/40) IgE-binding reactivity, whereas recombinant Der f 10 showed 25% (10/40) IgE-binding reactivity against the same sera from storage mite-sensitized and house dust mite-sensitized subjects. Both recombinant Tyr p 10 and Der f 10 showed little inhibition of IgE binding to T. putrescentiae crude extract by ELISA. Tropomyosin seems to contribute only a small portion of the cross-reactivity with house dust mites.
authors.affiliation: Department of Parasitology and Institute of Tropical Medicine, Brain Korea 21 Project for Medical Science, Yonsei University, College of Medicine, Seoul, Korea. jeongky@paran.com
authors.forename: Kyoung Yong
authors.initials: KY
authors.lastname: Jeong
authors.forename: Haeseok
authors.initials: H
authors.lastname: Lee
authors.forename: Jae Sik
authors.initials: JS
authors.lastname: Lee
authors.forename: Jongweon
authors.initials: J
authors.lastname: Lee
authors.forename: In-Yong
authors.initials: IY
authors.lastname: Lee
authors.forename: Han-Il
authors.initials: HI
authors.lastname: Ree
authors.forename: Chein-Soo
authors.initials: CS
authors.lastname: Hong
authors.forename: Tai-Soon
authors.initials: TS
authors.lastname: Yong
mesh_terms: D040041:Acaridae / Q000737:chemistry*; D000293:Adolescent; D000328:Adult; D000368:Aged; D000485:Allergens / Q000096:biosynthesis / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D002648:Child; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007073:Immunoglobulin E / Q000378:metabolism; D051379:Mice; D008875:Middle Aged; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000378:metabolism; D016415:Sequence Alignment; D014335:Tropomyosin / Q000096:biosynthesis / Q000276:immunology*
chemical_list: D000485:Allergens; D011994:Recombinant Proteins; D014335:Tropomyosin; C523282:Tyr p 10 allergen, Tyrophagus putrescentiae; D007073:Immunoglobulin E