Highly efficient green fluorescent protein-based kinase substrates.
Highly efficient green fluorescent protein-based kinase substrates.
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基于绿色荧光蛋白的高效激酶底物。
DOI：
10.1006/abio.1998.2885
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Shokat,KM
中科院分区：
文献类型：
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作者：
Yang,F;Liu,Y;Bixby,SD;Friedman,JD;Shokat,KM
我们已经开发了一种通用策略，通过将可磷酸化肽序列连接到his6标记的绿色荧光蛋白（GFP）的c端来设计高效的蛋白激酶蛋白底物。我们发现GFP中的几个c端连接位点允许将可磷酸化的尾部正确地呈现给各种蛋白激酶。利用这一策略，我们构建了高效的基于gfp的Src、C - abl、蛋白激酶A和蛋白激酶C βII蛋白激酶底物。Src的工程GFP底物（GFP235IYGEFG）比最常用的Src底物——兔肌烯醇酶的效率高300倍。
We have developed a general strategy for designing efficient protein substrates of protein kinases by attaching a phosphorylatable peptide sequence to the C-terminus of His6-tagged green fluorescent protein (GFP). We found that several C-terminal attachment sites in GFP allow for correct presentation of the phosphorylatable tail to a variety of protein kinases. Using this strategy, we have constructed highly efficient GFP-based substrates for Src, c-Abl, protein kinase A, and protein kinase C βII protein kinases. The engineered GFP substrate for Src (GFP235IYGEFG) is 300 times more efficient than the protein most commonly used as a Src substrate–rabbit muscle enolase.