Substrate specificity of protein tyrosine phosphatases 1B, RPTPα, SHP-1, and SHP-2.
Substrate specificity of protein tyrosine phosphatases 1B, RPTPα, SHP-1, and SHP-2.
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DOI：
10.1021/bi1014453
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发表时间：
2011-03-29
期刊：
影响因子：
2.9
通讯作者：
Pei, Dehua
中科院分区：
文献类型：
--
作者：
Ren, Lige;Chen, Xianwen;Luechapanichkul, Rinrada;Selner, Nicholas G.;Meyer, Tiffany M.;Wavreille, Anne-Sophie;Chan, Richard;Iorio, Caterina;Zhou, Xiang;Neel, Benjamin G.;Pei, Dehua
我们通过对蛋白质酪氨酸磷酸酶(PTP)α、PTP1B、SHP-1和SHP-2的组合多肽文库的珠状筛选和对单独合成的磷酸酪氨酰(Py)多肽的液相动力学分析，确定了PTP1B、RPTP、SHP-1和SHP-2的底物特异性。这些PTP表现出不同程度的序列特异性和催化效率。RPTPα的催化结构域具有非常弱的序列特异性，其活性大约比其他三个PTP低两个数量级。PTP1B催化结构域对Py两侧的酸性残基有一定的偏好，对多个磷酸化肽具有高度的活性，但不利于任何位置的碱性残基，Py−1位的Gly或Py+1位的Pro。相比之下，SHP-1和SHP-2有相似的底物特异性，但范围要窄得多，对Py残基两侧的酸性和芳香族疏水氨基酸有强烈的偏好。一个有效的SHP-1/2底物通常在N-末端含有两个或多个酸性残基，在Py的C-末端含有一个或多个酸性残基，但不含碱性残基。SHP-1和SHP-2之间存在细微的差异，因为SHP-1对Py−1和Py+1位的酸性残基有较强的偏好，而两种SHP对不同位置的酸性残基的偏好程度不同。对PTP1B、SHP-1和SHP-2的已知蛋白质底物的调查表明，体内去磷酸化模式与来自文库筛选的体外特异性图谱之间具有很好的一致性。这些结果表明，不同的PTP具有不同的序列专一性，PTP结构域的内在活性/专一性是决定该酶在体内底物专一性的重要因素。
We determined the substrate specificities of the protein tyrosine phosphatases (PTPs) PTP1B, RPTPα, SHP-1, and SHP-2 by on-bead screening of combinatorial peptide libraries and solution-phase kinetic analysis of individually synthesized phosphotyrosyl (pY) peptides. These PTPs exhibit different levels of sequence specificity and catalytic efficiency. The catalytic domain of RPTPα has very weak sequence specificity and is approximately two orders of magnitude less active than the other three PTPs. The PTP1B catalytic domain has modest preference for acidic residues on both sides of pY, is highly active towards multiply phosphorylated peptides, but disfavors basic residues at any position, a Gly at the pY−1 position, or a Pro at the pY+1 position. By contrast, SHP-1 and SHP-2 share similar but much narrower substrate specificities, with a strong preference for acidic and aromatic hydrophobic amino acids on both sides of the pY residue. An efficient SHP-1/2 substrate generally contains two or more acidic residues on the N-terminal side and one or more acidic residues on the C-terminal side of pY, but no basic residues. Subtle differences exist between SHP-1 and SHP-2 in that SHP-1 has a stronger preference for acidic residues at the pY−1 and pY+1 positions and the two SHPs prefer acidic residues at different positions N-terminal to pY. A survey of the known protein substrates of PTP1B, SHP-1, and SHP-2 shows an excellent agreement between the in vivo dephosphorylation pattern and the in vitro specificity profiles derived from library screening. These results suggest that different PTPs have distinct sequence specificity profiles and the intrinsic activity/specificity of the PTP domain is an important determinant of the enzyme’s in vivo substrate specificity.
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影响因子：
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