Purification and identification of an IgE suppressor from strawberry in an in vitro immunization system
Tóm tắt
We purified and identified an IgE suppressor from the strawberry ‘Toyonoka’, based on the decrease of IgE production in in vitro immunization (IVI). Gel filtration experiment indicated that fractions in a 15–48 kDa range and <10 kDa have an IgE suppressive activity. Furthermore, the fraction in 15–48 kDa was subjected to chromatofocusing and found to have activities at isoelectric points, pI 6.0, 7.0, and 8.0–9.2. We focused on the active fractions of pI 8.0–9.2 and the purified a large amount of strawberry extracts by cation exchange resins in batch. A purified 39 kDa protein showed homology to plant glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in N-terminal amino acid sequence and had GAPDH enzymatic activity. Nucleotide sequence and deduced amino acid sequence of the obtained cDNA clone of the protein matched with the sequence of Fragaria x ananassa GAPDH in the GenBank with >98% identical nucleotides and >99% identical amino acids, respectively. The purified strawberry GAPDH suppressed total IgE production in IVI in a dose-dependent manner. From these results, we identified GAPDH as IgE suppressor in the strawberry. Our study may be applicable to the development of new methods to relieve allergic conditions using GAPDH and the screening of other functional factors for human health.
Tài liệu tham khảo
Aaby K, Wrolstad RE, Ekeberg D, Skrede G (2007) Polyphenol composition and antioxidant activity in strawberry purees; impact of achene level and storage. J Agric Food Chem 55:5156–5166
Corry DB, Kheradmand F (1999) Induction and regulation of the IgE response. Nature 402:B18–B23
Ferdinand W (1964) The isolation and specific activity of rabbit muscle glyceraldehyde phosphate dehydrogenase. Biochem J 92:578
Hara MR, Cascio MB, Sawa A (2006) GAPDH as a sensor of NO stress. Biochim Biophys Acta 1762:502–509
Hassanain E, Silverberg JI, Norowitz KB, Chice S, Bluth MH, Brody N, Joks R, Durkin HG, Smith-Norowitz TA (2010) Green tea (Camelia sinensis) suppresses B cell production of IgE without inducing apoptosis. Ann Clin Lab Sci 40:135–143
Holgate ST (1999) The epidemic of allergy and asthma. Nature 402:B2–B4
Kawahara H, Maeda-Yamamoto M, Hakamata K (2000) Effective induction and acquisition of human monoclonal IgE antibodies reactive with house-dust mite extracts. J Immunol Methods 233:33–40
Kubo M, Inoue H (2006) Suppressor of cytokine signaling 3 (SOCS3) in Th2 cells evokes Th2 cytokines, IgE, and eosinophilia. Curr Allergy Asthma Rep 6:32–39
Metzger H, Alcaraz G, Hohman R, Kinet JP, Pribluda V, Quarto R (1986) The receptor with high affinity for immunoglobulin E. Annu Rev Immunol 4:419–470
Mitsuda K, Inoue A, Nishino N, Inoue Y, Kawahara H (2010) Detection of anti-allergic effects in strawberry extracts. In: Kamihira M et al. (eds) Proceedings of the 21st annual and international meeting of the Japanese association for animal cell technology, Fukuoka, November 2008. Animal cell technology: basic and applied aspects, vol 16. Springer, Dordrecht, The Netherlands, pp 353–358
Ninomiya M, Itoh T, Ishikawa S, Saiki M, Narumiya K, Yasuda M, Koshikawa K, Nozawa Y, Koketsu M (2010) Phenolic constituents isolated from Fragaria ananassa Duch inhibit antigen-stimulated degranulation through direct inhibition of spleen tyrosine kinase activation. Bioorg Med Chem 18:5932–5937
Sirover MA (1999) New insights into an old protein: the functional diversity of mammalian glyceraldehyde-3-phosphate dehydrogenase. Biochim Biophys Acta 1432:159–184
Sugahara T, Shirahata S, Sasaki T, Murakami H (1995) The mode of actions of glyceraldehyde-3-phosphate dehydrogenase identified as an immunoglobulin production stimulating factor. FEBS Lett 368:92–96
Yamaji R, Chatani E, Harada N, Sugimoto K, Inui H, Nakano Y (2005) Glyceraldehyde-3-phosphate dehydrogenase in the extracellular space inhibits cell spreading. Biochim Biophys Acta 1726:261–271
Yano S, Umeda D, Yamashita T, Ninomiya Y, Sumida M, Fujimura Y, Yamada K, Tachibana H (2007) Dietary flavones suppresses IgE and Th2 cytokines in OVA-immunized BALB/c mice. Eur J Nutr 46:257–263
Zhang Y, Seeram NP, Lee R, Feng L, Herber D (2008) Isolation and identification of strawberry phenolics with antioxidant and human cancer cell antiproliferative properties. J Agric Food Chem 56:670–675