通过斑点肽分析测定域介导的蛋白质-蛋白质相互作用。

Mapping of domain-mediated protein-protein interaction by SPOT peptide assay.

机构信息

University Medical Center Goettingen, 37077 Goettingen, Germany.

Ruhr University of Bochum, 44791 Bochum, Germany.

出版信息

STAR Protoc. 2021 Apr 27;2(2):100503. doi: 10.1016/j.xpro.2021.100503. eCollection 2021 Jun 18.

DOI:10.1016/j.xpro.2021.100503

PMID:33997817

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8100624/

Abstract

Identification of peptides mediating protein-protein interaction (PPI) is crucial for understanding the function of interlinked proteins in cellular processes and amino acid-associated diseases. Traditional PPI assays are laborious, involving the generation of many truncated proteins. SPOT peptide assay allows high-throughput detection of domains essential for PPI by synthesizing several hundred peptides on a cellulose membrane. Here, we present a rapid SPOT peptide protocol for identifying the binding motifs, which mediate interaction between the chromatin remodeling factors BAF155/BAF170 and the epigenetic factor Kdm6b. For complete details on the use and execution of this protocol, please refer to Narayanan et al. (2015).

摘要

鉴定介导蛋白质-蛋白质相互作用（PPI）的肽对于理解细胞过程中相互关联的蛋白质和与氨基酸相关的疾病中的功能至关重要。传统的 PPI 测定方法繁琐，涉及生成许多截断的蛋白质。SPOT 肽测定法通过在纤维素膜上合成数百种肽来允许高通量检测 PPI 必需的结构域。在这里，我们提出了一种快速的 SPOT 肽方案，用于鉴定介导染色质重塑因子 BAF155/BAF170 和表观遗传因子 Kdm6b 之间相互作用的结合基序。有关此方案的使用和执行的完整详细信息，请参阅 Narayanan 等人。（2015 年）。