One-step purification of the beta-glucan elicitor-binding protein from soybean (Glycine max L) roots and characterization of an anti-peptide antiserum

被引:55
作者
Mithofer, A
Lottspeich, F
Ebel, J
机构
[1] UNIV MUNICH,INST BOT,D-80638 MUNICH,GERMANY
[2] MAX PLANCK INST BIOCHEM,D-82152 MARTINSRIED,GERMANY
关键词
beta-glucan-binding protein; glucan affinity purification; peptide-specific antiserum; soybean;
D O I
10.1016/0014-5793(96)00126-3
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A low abundance beta-glucan elicitor-binding protein from soybean was isolated by a rapid, simple and one-step purification method yielding about 9000-fold enrichment, The affinity-based purification technique was more efficient than a procedure that uses conventional methods and preserved the binding activity to a much larger extent, The final preparation consisted of one major protein with an apparent molecular mass of about 75 kDa, Electrophoretic analyses of the purified and photoaffinity-labeled binding protein showed that the native protein was an oligomer with apparent molecular mass of about 240 kDa, A polyclonal anti-peptide antiserum was raised against a synthetic 15-mer internal oligopeptide sequence derived from the 75-kDa protein, The antiserum recognized the purified binding protein in immunoblotting experiments and precipitated the affinity-labeled protein from a crude extract of the membrane fraction.
引用
收藏
页码:203 / 207
页数:5
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