NONHYDROLYZABLE PHOSPHOTYROSYL MIMETICS FOR THE PREPARATION OF PHOSPHATASE-RESISTANT SH2 DOMAIN INHIBITORS
NONHYDROLYZABLE PHOSPHOTYROSYL MIMETICS FOR THE PREPARATION OF PHOSPHATASE-RESISTANT SH2 DOMAIN INHIBITORS
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DOI：
10.1021/bi00187a015
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发表时间：
1994-05-31
期刊：
影响因子：
2.9
通讯作者：
SHOELSON, SE
中科院分区：
文献类型：
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作者：
BURKE, TR;SMYTH, MS;SHOELSON, SE
Src同源性2 （SH2）结构域通过高亲和力结合磷酸酪氨酸（pTyr）承载蛋白序列，参与蛋白酪氨酸激酶（PTK）介导的细胞信号转导。虽然含有pTyr的多肽以序列特异性的方式竞争性地抑制磷酸化蛋白和同源SH2蛋白之间的结合，但这种多肽会被细胞磷酸酶迅速去磷酸化。我们现在描述我们的努力开发SR2抑制肽含有磷酸酶抗性pTyr替代物。母体化合物（磷甲酰乙基）苯丙氨酸（Pmp）是一种基于磷酸盐的pTyr的模拟物，其中磷酸酯氧（>COPO3H2）被亚甲基单元(>CCX(2)PO(3)H（2)， X(2) = H-2）取代，Pmp类似物在膦酸-亚甲基碳上含有氟(X（2) = H， F或X(2) = F-2）或羟基（X(2) = H， OH）取代基，并被纳入肽中用作SH2结构域抑制剂。在一项利用磷脂酰肌醇（PI） 3-激酶的c端SH2结构域进行的分析中，具有GXVPML序列的肽[其中X = pTyr, Pmp，羟基Pmp (HPmp)，单氟Pmp （FPmp）和双氟Pmp (F(2)Pmp)]显示出的结合能力顺序为HPmp < Pmp < FPmp < F(2)Pmp = pTyr。用pTyr和F(2)Pmp制备了选择性结合Src和Grb2 SH2结构域的不同肽序列。与类似的pTyr肽相比，F(2)Pmp肽结合的相对亲和力高（0.2- 5倍）。我们得出结论，含有F(2)Pmp的肽以高亲和力和特异性结合SH2结构域，并且对细胞磷酸酶具有抗性，应该为破坏完整细胞中SH2结构域介导的信号通路提供一种普遍有用的工具。
Src homology 2 (SH2) domains participate in protein tyrosine kinase (PTK)-mediated cellular signal transduction through their ability to bind with high affinity to phosphotyrosyl (pTyr)-bearing protein sequences. Although peptides containing pTyr competitively inhibit the binding between phosphoproteins and cognate SH2 proteins in a sequence-specific manner, such peptides are rapidly dephosphorylated by cellular phosphatases. We now describe our efforts to develop SR2 inhibitory peptides containing phosphatase-resistant pTyr Surrogates. The parent compound, (phosphonomethyl)phenylalanine (Pmp),is a phosphonate-based mimetic of pTyr in which the phosphate ester oxygen (>COPO3H2) has been replaced by a methylene unit (>CCX(2)PO(3)H(2), X(2) = H-2) Pmp analogues bearing fluorine (X(2) = H, F or X(2) = F-2) or hydroxyl (X(2) = H, OH) substituents on the phosphonate alpha-methylene carbon have been prepared and incorporated into peptides for use as SH2 domain inhibitors. In an assay using the C-terminal SH2 domain of phosphatidylinositol (PI) 3-kinase, peptides having a GXVPML sequence [where X = pTyr, Pmp, hydroxy-Pmp (HPmp), monofluoro-Pmp (FPmp), and difluoro-Pmp (F(2)Pmp)] exhibited binding potency in the order HPmp < Pmp < FPmp < F(2)Pmp = pTyr. Distinct peptide sequences which bind selectively with Src and Grb2 SH2 domains were also prepared with pTyr and F(2)Pmp. The F(2)Pmp peptides bound with high (0.2- to 5-fold) relative affinity, compared to analogous pTyr peptides. We conclude that peptides containing F(2)Pmp bind to SH2 domains with high affinity and specificity and, being resistant to cellular phosphatases, should provide a generally useful tool for disrupting SH2 domain-mediated signaling pathways in intact cells.