A 33-kDa allergen from rice (Oryza sativa L. Japonica) -: cDNA cloning, expression, and identification as a novel glyoxalase I
A 33-kDa allergen from rice (Oryza sativa L. Japonica) -: cDNA cloning, expression, and identification as a novel glyoxalase I
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DOI：
10.1074/jbc.m010337200
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发表时间：
2001-04-06
影响因子：
4.8
通讯作者：
Matsuda, T
中科院分区：
文献类型：
--
作者：
Usui, Y;Nakase, M;Matsuda, T
众所周知，谷类蛋白会引起过敏反应，如某些人群的贝克氏哮喘和严重的特应性皮炎。在大米过敏中，分子质量为14-16、26、33和56 kDa的蛋白质已被证明是潜在的过敏原。在本研究中，为了鉴定和鉴定33 kDa变应原，命名为Glb33，首先将该蛋白纯化为均一，并分离其cDNA克隆。在大肠杆菌中表达的重组Glb33与天然Glb33具有与小鼠免疫球蛋白和患者抗Glb33免疫球蛋白的反应能力。Glb33基因编码291个氨基酸，有两个120个氨基酸残基重复序列，氨基酸序列与人、植物、酵母和细菌等各种生物的乙草酸酶I序列相似。不出所料，从水稻种子中提纯的天然Glb33和重组蛋白都具有催化甲基乙醛和谷胱甘肽缩合成S-乳糖基谷胱甘肽的乙醛酶I活性。然而，Glb33与细菌乙二醛酶I有较高的序列同源性，而不是与已知的植物和酵母酶。Glb33转录本和蛋白不仅在成熟的水稻种子中检测到，而且在水稻的茎和叶中也检测到。综上所述，水稻过敏原Glb33被鉴定为一种新型的植物乙二醛酶I，在包括成熟种子在内的各种植物组织中都有表达。
Cereal proteins are known to cause allergic reactions such as Baker's asthma and severe atopic dermatitis to certain populations. In rice allergy, proteins with molecular masses of 14-16, 26, 33, and 56 kDa have been demonstrated to be potentially allergenic. In this study, to identify and characterize the 33-kDa allergen, designated Glb33, this protein was first purified to homogeneity, and its cDNA clone was isolated. When expressed in Escherichia coli, the recombinant Glb33 was shown to be as reactive as the native Glb33 with mouse IgG and patients' IgE antibodies to Glb33. The Glb33 cDNA coded for a protein of 291 amino acids with two 120-amino acid residue repeats, and the amino acid sequence showed similarity to glyoxalase I from various organisms, including human, plant, yeast, and bacterium. As expected, both native Glb33 purified from rice seeds and the recombinant protein had glyoxalase I activity that catalyzes condensation of methylglyoxal and glutathione into S-lactoylglutathione. However, Glb33 had a higher sequence identity to the bacterial glyoxalase I rather than to known plant and yeast enzymes. Both the Glb33 transcript and the protein were detected not only in maturing seeds of rice but also in its stem and leaf. Taken all together, the rice allergen, Glb33, was identified to be a novel type of plant glyoxalase I that is expressed in various plant tissues, including maturing seeds.