Yin Jun, Liu Fei, Li Xiaohua, Walsh Christopher T
Department of Biological Chemistry and Molecular Pharmacology and Institute of Chemistry and Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, Massachusetts 02115, USA.
J Am Chem Soc. 2004 Jun 30;126(25):7754-5. doi: 10.1021/ja047749k.
We report here the development of a general strategy for site-specific labeling of proteins with small molecules by posttranslational modification enzyme, phosphopantetheinyl transferase Sfp. The target proteins are expressed as fusions to the peptide carrier protein (PCP) excised from nonribosomal peptide synthetase, and Sfp catalyzes the covalent modification of a specific serine residue on PCP by the small molecule-phosphopantetheinyl conjugate. The labeling reaction proceeds with high specificity and efficiency, targeting PCP fusion proteins in the cell lysate. The PCP tag has been shown to be compatible with various proteins, and Sfp-catalyzed PCP modification, compatible with various small-molecule probes conjugated to coenzyme A, highlighting the potential of the PCP tag for site-specific protein labeling with small molecules.
我们在此报告了一种通用策略的开发，该策略通过翻译后修饰酶磷酸泛酰巯基乙胺基转移酶Sfp对蛋白质进行小分子位点特异性标记。目标蛋白表达为与从非核糖体肽合成酶中切除的肽载体蛋白（PCP）的融合蛋白，并且Sfp催化小分子-磷酸泛酰巯基乙胺共轭物对PCP上特定丝氨酸残基的共价修饰。标记反应以高特异性和高效率进行，靶向细胞裂解物中的PCP融合蛋白。已证明PCP标签与各种蛋白质兼容，并且Sfp催化的PCP修饰与与辅酶A缀合的各种小分子探针兼容，突出了PCP标签用于小分子位点特异性蛋白质标记的潜力。
