Protein-protein interaction detection in vitro and in cells by proximity biotinylation.
作者： Marta Fernández-Suárez , T. Scott Chen , Alice Y. Ting
DOI: 10.1021/JA801445P
关键词:
摘要: We report a new method for detection of protein−protein interactions in vitro and cells. One protein partner is fused to Escherichia coli biotin ligase (BirA), while the other BirA’s “acceptor peptide” (AP) substrate. If two proteins interact, BirA will catalyze site-specific biotinylation AP, which can be detected by streptavidin staining. To minimize nonspecific signals, we engineered AP sequence reduce its intrinsic affinity BirA. The rapamycin-controlled interaction between FKBP FRB could cells with signal background ratio as high 28. also extended imaging phosphorylation-dependent Cdc25C phosphatase 14-3-3e phosphoserine/threonine binding protein. Protein−protein proximity has advantages low background, sensitivity, small tag size, good spatial resolution
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