Onishi Y, Kuroda M, Yasueda H, Saito A, Sono-Koyama E, Tunasawa S, Hashida-Okado T, Yagihara T, Uchida K, Yamaguchi H, Akiyama K, Kato I, Takesako K
Biotechnology Research Laboratories, Takara Shuzo Co. Ltd, Shiga, Japan.
Eur J Biochem. 1999 Apr;261(1):148-54. doi: 10.1046/j.1432-1327.1999.00247.x.
The yeast Malassezia furfur is a natural inhabitant of the human skin microflora that induces an allergic reaction in atopic dermatitis. To identify allergens of M. furfur, we separated a crude preparation of M. furfur antigens as discrete spots by 2-D PAGE and detected IgE-binding proteins using sera of atopic dermatitis patients. We identified the known allergens, Mal f 2 and Mal f 3, and determined N-terminal amino acid sequences of six new IgE-binding proteins including Mal f 4. The cDNA and genomic DNA encoding Mal f 4 were cloned and sequenced. The gene was mitochondrial malate dehydrogenase and encoded Mal f 4 composed of 315 amino acids and a signal sequence of 27 amino acids. We purified Mal f 4, which had a molecular mass of 35 kDa from a membrane fraction of a lysate of cultured cells. Thirty of 36 M. furfur-allergic atopic dermatitis patients (83.3%) had elevated serum levels of IgE to purified Mal f 4, indicating that Mal f 4 is a major allergen. There was a significant correlation of the Phadebas RAST unit values of Mal f 4 and the crude antigen, but not between Mal f 4 and the known allergen Mal f 2.
糠秕马拉色菌是人类皮肤微生物群的天然寄居菌，可在特应性皮炎中引发过敏反应。为了鉴定糠秕马拉色菌的变应原，我们通过二维聚丙烯酰胺凝胶电泳将糠秕马拉色菌抗原粗制品分离为离散斑点，并使用特应性皮炎患者的血清检测免疫球蛋白E结合蛋白。我们鉴定出了已知变应原Mal f 2和Mal f 3，并确定了包括Mal f 4在内的6种新的免疫球蛋白E结合蛋白的N端氨基酸序列。对编码Mal f 4的cDNA和基因组DNA进行了克隆和测序。该基因是线粒体苹果酸脱氢酶，编码由315个氨基酸组成的Mal f 4和一个27个氨基酸的信号序列。我们从培养细胞裂解物的膜部分纯化了分子量为35 kDa的Mal f 4。36名对糠秕马拉色菌过敏的特应性皮炎患者中有30名（83.3%）血清中免疫球蛋白E对纯化的Mal f 4水平升高，表明Mal f 4是主要变应原。Mal f 4与粗抗原的Phadebas RAST单位值之间存在显著相关性，但Mal f 4与已知变应原Mal f 2之间不存在显著相关性。
