通过蓝色天然聚丙烯酰胺凝胶电泳阻滞分析法检测蛋白质复合物相互作用。

Detection of protein complex interactions via a Blue Native-PAGE retardation assay.

作者信息

Swamy Mahima, Molnar Eszter, Bock Thomas, Bausch-Fluck Damaris, Wollscheid Bernd, Schamel Wolfgang W A

机构信息

Department for Molecular Immunology, Max Planck-Institute for Immunobiology, 79108 Freiburg, Germany.

出版信息

Anal Biochem. 2009 Sep 15;392(2):177-9. doi: 10.1016/j.ab.2009.05.039. Epub 2009 May 29.

DOI:10.1016/j.ab.2009.05.039

PMID:19482002

Abstract

We describe the Blue Native (BN)-PAGE retardation assay for the detection of interactions of biomolecules with protein complexes. Potential interactors of proteins are included in the BN gel matrix, resulting in retardation of proteins that interact with the added molecule. After validation using the T-cell antigen receptor, we applied the assay for a general identification of dextran interactors in combination with mass spectroscopy. The proteomic screen revealed triosephosphate isomerase oligomer as a dextran-binding, high M(R) complex.

摘要

我们描述了用于检测生物分子与蛋白质复合物相互作用的蓝色原胶（BN）-聚丙烯酰胺凝胶电泳阻滞分析方法。蛋白质的潜在相互作用分子被包含在BN凝胶基质中，导致与添加分子相互作用的蛋白质迁移受阻。在用T细胞抗原受体进行验证后，我们将该分析方法应用于结合质谱法对葡聚糖相互作用分子进行全面鉴定。蛋白质组学筛选揭示了磷酸丙糖异构酶寡聚体是一种与葡聚糖结合的高分子量复合物。