茄病镰刀菌65kd过敏原Fus sI3596*的纯化与鉴定

Purification and characterization of Fus sI3596*, a 65 kd allergen of Fusarium solani.

作者信息

Verma J, Pasha S, Gangal S V

机构信息

Centre for Biochemical Technology, Delhi, India.

出版信息

Mol Cell Biochem. 1994 Feb 23;131(2):157-66. doi: 10.1007/BF00925952.

DOI:10.1007/BF00925952

PMID:8035781

Abstract

A component of Fusarium solani (F. solani), identified as the major allergen, Fus sI3596* was purified to homogeneity from culture filtrate (CF) by means of anion-exchange column chromatography, gel filtration and FPLC. The homogeneity of Fus sI3596* was assessed by IEF, PAGE, SDS-PAGE (non-reducing), immunoblot and HPLC. Fus sI3596* was isolated as a glycoprotein of MW 65 kd and pI 3.6. The IgE ELISA-inhibition assay after periodate treatment of the fraction showed a lower IgE binding capacity suggesting involvement of carbohydrate moiety in IgE binding reactions of the allergen. Peptide fragments of Fus sI3596* obtained after CNBr and trypsin treatment were analysed by immunoblotting for their allergenicity. This study indicated that there could be at least 3 allergenic determinants in the major allergen, Fus sI3596* of F. solani CF.

摘要

茄病镰刀菌（F. solani）的一种成分，被鉴定为主要过敏原Fus sI3596*，通过阴离子交换柱色谱、凝胶过滤和快速蛋白质液相色谱（FPLC）从培养滤液（CF）中纯化至同质。通过等电聚焦（IEF）、聚丙烯酰胺凝胶电泳（PAGE）、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE，非还原）、免疫印迹和高效液相色谱（HPLC）评估Fus sI3596的同质性。Fus sI3596被分离为一种分子量65千道尔顿、等电点3.6的糖蛋白。对该级分进行高碘酸盐处理后的IgE酶联免疫吸附测定抑制试验显示，IgE结合能力较低，表明碳水化合物部分参与了过敏原的IgE结合反应。通过免疫印迹分析经溴化氰（CNBr）和胰蛋白酶处理后获得的Fus sI3596的肽片段的致敏性。本研究表明，茄病镰刀菌CF的主要过敏原Fus sI3596中可能至少存在3个致敏决定簇。