Expression of tropomyosin from Blattella germanica as a recombinant non-fusion protein in Pichia pastoris and comparison of its IgE reactivity with its native counterpart

Protein Expression and Purification
Kyoung Yong JeongTai-Soon Yong

Abstract

Tropomyosins derived from invertebrates are well-known pan allergens. However, the allergenicities of recombinant tropomyosins are variable. Here, we undertook to compare the IgE-binding reactivities of native and recombinant German cockroach tropomyosins. Native tropomyosin was purified by ammonium sulfate fractionation, hydroxyapatite column chromatography, and electroelution, and recombinant tropomyosin was expressed in Pichia pastoris. The allergenicities of the native and recombinant tropomyosins were compared by ELISA inhibition analysis. Native German cockroach tropomyosin showed 18% IgE-binding reactivity to German cockroach sensitized sera. Recombinant tropomyosin was produced without fusion protein and its N-terminus was blocked like that of the native counterpart. The IgE-binding reactivity of the recombinant was found to be comparable to that of native tropomyosin over the concentration range 1-1000 ng/ml by ELISA inhibition testing. Recombinant German cockroach tropomyosin expressed in Pichia pastoris showed better allergenicity than that expressed in Escherichia coli. Other factors in addition to the structural differences of native and recombinant proteins may also influence the IgE reactivities of tropomyosins.

References

Mar 1, 1991·The Journal of Allergy and Clinical Immunology·T A Platts-Mills, M D Chapman
Mar 1, 1972·Canadian Journal of Biochemistry·R S Hodges, L B Smillie
Jan 1, 1982·Methods in Enzymology·L B Smillie
May 1, 1995·Allergy·O Scheiner, D Kraft
May 1, 1995·International Archives of Allergy and Immunology·L K ArrudaM D Chapman
Aug 1, 1996·The Journal of Allergy and Clinical Immunology·H S NelsonA Buchmeier
May 30, 1998·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·R SluyterW J Britton
Sep 4, 1999·International Archives of Allergy and Immunology·G ReeseS B Lehrer
Jan 21, 2000·FEMS Microbiology Reviews·J L Cereghino, J M Cregg
Feb 25, 2000·International Archives of Allergy and Immunology·P ParronchiE Maggi
Sep 26, 2000·Allergy·J A AsturiasA Martínez
Feb 13, 2001·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·T YasuharaY Okumura
Apr 9, 2001·Immunological Reviews·R Valenta, D Kraft
Jun 21, 2001·Biochemical and Biophysical Research Communications·E HilarioM C Bertolini
Sep 12, 2001·Allergy·S G JohanssonUNKNOWN EAACI (the European Academy of Allergology and Cinical Immunology) nomenclature task force
Jan 16, 2002·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Reinhard HillerRudolf Valenta
Jan 31, 2002·American Journal of Respiratory and Critical Care Medicine·Anna PomésVenugopal Dhanaraj
Apr 20, 2002·Current Opinion in Allergy and Clinical Immunology·A Pomés, M D Chapman
May 8, 2002·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·M L Patterson, J E Slater
May 22, 2002·International Archives of Allergy and Immunology·Lili Kazemi-ShiraziRudolf Valenta
Jul 18, 2002·International Archives of Allergy and Immunology·Fátima FerreiraChristof Ebner
Aug 23, 2002·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·F C YiB W Lee
Jan 22, 2003·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·C HarwaneggR Valenta
Jun 25, 2003·Protein Expression and Purification·Marcos Castanheira AlegriaMaria Célia Bertolini

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Citations

Aug 11, 2005·Current Allergy and Asthma Reports·L Karla Arruda
Jan 27, 2010·Developmental and Comparative Immunology·Beatrice MindykowskiHeinz Decker
Jan 13, 2009·Biomedical and Environmental Sciences : BES·Chuan WangXiao-Fang Pei
Nov 4, 2009·The Korean Journal of Parasitology·Tai-Soon Yong, Kyoung Yong Jeong
Jul 1, 2005·Physiological Reviews·Scott L Hooper, Jeffrey B Thuma

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