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

FEBS Letters

Lottspeich, Friedrich²  Ebel, Jürgen¹  Mithöfer, Axel¹ 

[1] Botanisches Institut der Universität, Menzinger Str. 67, D-80638 München, Germany;Max-Planck-Institut für Biochemie, D-82152 Martinsried, Germany

关键词: β-Glucan-binding protein;    Glucan affinity purification;    Peptide-specific antiserum;    Soybean;   

DOI  :  10.1016/0014-5793(96)00126-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A low abundance β-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.

【 授权许可】

Unknown