Background: In spite of being among the main foods responsible for allergic reactions worldwide, soybean (Glycine max)-derived products continue to be increasingly widespread in a variety of food products due to their well-documented health benefits. Soybean also continues to be one of the elected target crops for genetic modification. The aim of this study was to characterize the soya proteome and, specifically, IgE-reactive proteins as well as to compare the IgE response in soya-allergic individuals to genetically modified Roundup Ready soya® versus its non-transgenic control. Methods: We performed two-dimensional gel electrophoresis of protein extracts from a 5% genetically modified Roundup Ready flour sample and its non-transgenic control followed by Western blotting with plasma from 5 soya-sensitive individuals. We used peptide tandem mass spectrometry to identify soya proteins (55 protein matches), specifically IgE-binding ones, and to evaluate differences between transgenic and non-transgenic samples. Results: We identified 2 new potential soybean allergens – one is maturation associated and seems to be part of the late embryogenesis abundant proteins group and the other is a cysteine proteinase inhibitor. None of the individuals tested reacted differentially to the transgenic versus non-transgenic samples under study. Conclusion: Soybean endogenous allergen expression does not seem to be altered after genetic modification. Proteomics should be considered a powerful tool for functional characterization of plants and for food safety assessment.

1.
Taylor SL, Lehrer SB: Principles and characteristics of food allergens. Crit Rev Food Sci Nutr 1996;36(suppl):S91–S118.
2.
Helm RM, Burks AW: Mechanisms of food allergy. Curr Opin Immunol 2000;12:647–653.
3.
Food and Agriculture Organization of the United Nations: Report of the FAO Technical Consultation on Food Allergies. Rome, Food and Agriculture Organization of the United Nations, 1995.
4.
Herbert B: Advances in protein solubilization for two-dimensional electrophoresis. Electrophoresis 1999;20:660–663.
5.
Pandey A, Mann M: Proteomics to study genes and genomes. Nature 2000;405:837–846.
6.
Magni C, Ballabio C, Restani P, Sironi E, Scarafoni A, Poiesi C, Duranti M: Two-dimensional electrophoresis and Western-blotting analyses with anti Ara h 3 basic subunit IgG evidence the cross-reacting polypeptides of Arachis hypogaea, Glycine max, and Lupinus albus seed proteomes. J Agric Food Chem 2005;53:2275–2281.
7.
Herman EM, Helm RM, Jung R, Kinney AJ: Genetic modification removes an immunodominant allergen from soybean. Plant Physiol 2003;132:36–43.
8.
Hajduch M, Ganapathy A, Stein JW, Thelen JJ: A systematic proteomic study of seed filling in soybean. Establishment of high-resolution two-dimensional reference maps, expression profiles, and an interactive proteome database. Plant Physiol 2005;137:1397–1419.
9.
Mooney BP, Thelen JJ: High-throughput peptide mass fingerprinting of soybean seed proteins: automated workflow and utility of UniGene expressed sequence tag databases for protein identification. Phytochemistry 2004;65:1733–1744.
10.
Natarajan S, Xu C, Caperna TJ, Garret WM: Comparison of protein solubilization methods suitable for proteomic analysis of soybean seed proteins. Anal Biochem 2005;342:214–220.
11.
Ramagli LS: Quantifying protein in 2D-PAGE solubilization buffers. Methods Mol Biol 1999;112:99–103.
12.
Neuhoff V, Stamm R, Eibl H: Clear background and highly sensitive protein staining with Coomassie Blue dyes in polyacrylamide gels: a systematic analysis. Electrophoresis 1985;6:427–448.
13.
Pandey A, Anderson JS, Mann M: Use of mass spectrometry to study signaling pathways. Sci STKE 2000;37:PL1.
14.
Axén R, Drevin H, Kober A, Yman L: A new laboratory diagnostic system applied to allergy testing. N Engl Reg Allergy Proc 1988;9:503.
15.
Gatlin CL, Eng JK, Cross ST, Detter JC, Yater JR 3rd: Automated identification of amino acid sequence variations in proteins by HPLC/microspray tandem mass spectrometry. Anal Chem 2000;72:757–763.
16.
Perkins DN, Pappinl DJC, Creasy DM, Cottrell JS: Probability-based protein identification by searching databases using mass spectrometry data. Electrophoresis 1999;20:3551–3567.
17.
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227:680–685.
18.
Blum H, Beier H, Gross HJ: Improved silver staining of plant-proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 1987;8:93–99.
19.
Yaklich R, Helm R, Herman E: Analysis of the distribution of the major soybean allergen in a core collection of Glycine max accessions. Crop Sci 1999;39:1444–1447.
20.
Helm RM, Cockrell G, Herman E, Burks AW, Sampson HA, et al: Cellular and molecular characterization of a major soybean allergen. Int Arch Allergy Immunol 1998;117:29–37.
21.
Ali-Benali MA, Alary R, Joudrier P, Gautier M-F: Comparative expression of five Lea genes during wheat seed development and in response to abiotic stresses by real-time quantitative RT-PCR. Biochim Biophys Acta 2005;1730:56–65.
22.
Koiwa H, Shade RE, Zhu-Salzman K, D’Urzo MP, Murdock LL, et al: A plant defensive cystatin (soyacystatin) targets cathepsin L-like digestive cysteine proteinases (DvCALs) in the larval midgut of western corn rootworm (Diabrotica virgifera virgifera). FEBS Lett 2000;471:67–70.
23.
Arai S, Matsumoto I, Emori Y, Abe K: Plant seed cystatins and their target enzymes of endogenous origin. J Agric Food Chem 2002;50:6612–6617.
24.
Massonneau A, Condamine P, Wisniewski J-P, Zivy M,Rogowsky PM: Maize cystatins respond to developmental cues, cold stress and drought.Biochim Biophys Acta 2005;1729:186–199.
25.
Midoro-Horiuti T, Brooks EG, Goldblum RM: Pathogenesis-related proteins of plants as allergens. Ann Allergy Asthma Immunol 2001;87:261–271.
26.
Hoffmann-Sommergruber K: Pathogenesis-related (PR)-proteins identified as allergens. Biochem Soc Trans 2002;30:930–935.
27.
Corpillo D, Gardini G, Vaira AM, Basso M, Aime S, et al: Proteomics as a tool to improve investigation of substantial equivalence in genetically modified organisms: the case of a virus-resistant tomato. Proteomics 2004;4:193–200.
28.
Lehesranta SJ, Davies HV, Shepherd LVT, Nunan N, McNicol JW, et al: Comparison of tuber proteomes of potato varieties, landraces, and genetically modified lines. Plant Physiol 2005;138:1690–1699.
29.
Batista R, Nunes B, Carmo M, Cardoso C, José HS, et al: Lack of detectable allergenicity of transgenic maize and soya samples. J Allergy Clin Immunol 2005;116:403–410.
30.
Metcalfe DD: Genetically modified crops and allergenicity. Nat Immunol 2005;6:857–860.
31.
Taylor SL: Assessment of the allergenicity of genetically modified foods. Nutr Abstracts Rev (Series A) 1997;67:1163–1168.
32.
Mendieta NLR, Nagy A-M, Lints FA: The potential allergenicity of novel foods. J Sci Food Agric 1997;75:405–411.
33.
Gonzalez R, Polo F, Zapatero L, Caravaca F, Carreira J: Purification and characterization of major inhalant allergens from soybean hulls. Clin Exp Allergy1992;22:748–755.
34.
Gonzalez R, Varela J, Carreira J, Polo F: Soybean hydrophobic protein and soybean hull allergy. Lancet 1995;346:48–49.
35.
Codina R, Lockey RF, Fernandez-Caldas E, Rama R: Purification and characterization of a soybean hull allergen responsible for the Barcelona asthma outbreaks. 2. Purification and sequencing of the Gly m 2 allergen. Clin Exp Allergy 1997;27:424–430.
36.
Gonzalez R, Zapatero L, Caravaca F, Carreira J: Identification of soybean proteins responsible for respiratory allergies. Int Arch Allergy Appl Immunol 1991;95:53–57.
37.
Rihs HP, Chen Z, Rueff F, Petersen A, Rozynek P, et al: IgE binding of the recombinant allergen soybean profilin (rGly m 3) is mediated by conformational epitopes. J Allergy Clin Immunol 1999;104:1293–1301.
38.
Mittag D, Vieths S, Vogel L, Becker WM, Rihs HP, et al: Soybean allergy in patients allergic to birch pollen: clinical investigation and molecular characterization of allergens. J Allergy Clin Immunol 2004;113:148–154.
39.
Crowell DN, John ME, Russell D, Amasino RM: Characterization of a stress-induced, developmentally regulated gene family from soybean. Plant Mol Biol 1992;18:459–466.
40.
Kleine-Tebbe J, Vogel L, Crowell DN, Haustein UF, Vieths S: Severe oral allergy syndrome and anaphylactic reactions caused by a Bet v 1-related PR-10 protein in soybean, SAM22. J Allergy Clin Immunol 2002;110:797–804.
41.
Ogawa T, Bando N, Tsuji H, Okajima H, Nishikawa K, et al: Investigation of the IgE-binding proteins in soybeans by immunoblotting with the sera of the soybean-sensitive patients with atopic dermatitis. J Nutr Sci Vitaminol (Tokyo) 1991;37:555–565.
42.
Ogawa T, Bando N, Tsuji H, Nishikawa K, Kitamura K: Identification of the soybean allergenic protein, Gly m Bd 30K, with the soybean seed 34-kDa oil-body-associated protein. Biosci Biotechnol Biochem 1993;57:1030–1033.
43.
Tsuji H, Hiemori M, Kimoto M, Yamashita H, Kobatake R, et al: Cloning of cDNA encoding a soybean allergen, Gly m Bd 28K. Biochim Biophys Acta 2001;1518:178–182.
44.
Djurtoft R, Pedersen HS, Aabin B, Barkholt V: Studies of food allergens: soybean and egg proteins. Adv Exp Med Biol 1991;289:281–293.
45.
Gu X, Beardslee T, Zeece M, Sarath G, Markwell J: Identification of IgE-binding proteins in soy lecithin. Int Arch Allergy Immunol 2001;126:218–225.
46.
Lin J, Shewry PR, Archer DB, Beyer K, Niggemann B, et al: The potential allergenicity of two 2S albumins from soybean (Glycine max): a protein microarray approach. Int Arch Allergy Immunol 2006;141:91–102.
47.
Barnett D, Howden ME: Lectins and the radioallergosorbent test. J Allergy Clin Immunol 1987;80:558–561.
48.
Baur X, Pau M, Czuppon A, Fruhmann G: Characterization of soybean allergens causing sensitization of occupationally exposed bakers. Allergy 1996;51:326–330.
49.
Moroz LA, Yang WH: Kunitz soybean trypsin inhibitor: a specific allergen in food anaphylaxis. N Engl J Med 1980;302:1126–1128.
50.
Burks AW, Cockrell G, Connaughton C, Guin J, Allen W, et al: Identification of peanut agglutinin and soybean trypsin inhibitor as minor legume allergens. Int Arch Allergy Immunol 1994;105:143–149.
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.