抗生物素蛋白抗体可实现对蛋白质上生物素化位点的大规模检测。
Antibodies to biotin enable large-scale detection of biotinylation sites on proteins.
作者信息
Udeshi Namrata D, Pedram Kayvon, Svinkina Tanya, Fereshetian Shaunt, Myers Samuel A, Aygun Ozan, Krug Karsten, Clauser Karl, Ryan Dominic, Ast Tslil, Mootha Vamsi K, Ting Alice Y, Carr Steven A
机构信息
Proteomics, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Departments of Genetics, Biology, and Chemistry, Stanford University, Stanford, California, USA.
出版信息
Nat Methods. 2017 Dec;14(12):1167-1170. doi: 10.1038/nmeth.4465. Epub 2017 Oct 16.
Although purification of biotinylated molecules is highly efficient, identifying specific sites of biotinylation remains challenging. We show that anti-biotin antibodies enable unprecedented enrichment of biotinylated peptides from complex peptide mixtures. Live-cell proximity labeling using APEX peroxidase followed by anti-biotin enrichment and mass spectrometry yielded over 1,600 biotinylation sites on hundreds of proteins, an increase of more than 30-fold in the number of biotinylation sites identified compared to streptavidin-based enrichment of proteins.
尽管生物素化分子的纯化效率很高，但确定生物素化的特定位点仍然具有挑战性。我们表明，抗生物素抗体能够从复杂的肽混合物中以前所未有的方式富集生物素化肽。使用APEX过氧化物酶进行活细胞邻近标记，然后进行抗生物素富集和质谱分析，在数百种蛋白质上产生了超过1600个生物素化位点，与基于链霉亲和素的蛋白质富集相比，所鉴定的生物素化位点数量增加了30多倍。
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