...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Molecular and Cellular Biology >Structure-function analysis of SH3 domains: SH3 binding specificity altered by single amino acid substitutions.
【24h】

Structure-function analysis of SH3 domains: SH3 binding specificity altered by single amino acid substitutions.

机译：SH3结构域的结构功能分析：SH3结合特异性被单个氨基酸取代改变。

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

SH3 domains mediate intracellular protein-protein interactions through the recognition of proline-rich sequence motifs on cellular proteins. Structural analysis of the Src SH3 domain (Src SH3) complexed with proline-rich peptide ligands revealed three binding sites involved in this interaction: two hydrophobic interactions (between aliphatic proline dipeptides in the SH3 ligand and highly conserved aromatic residues on the surface of the SH3 domain), and one salt bridge (between Asp-99 of Src and an Arg three residues upstream of the conserved Pro-X-X-Pro motif in the ligand). We examined the importance of the arginine binding site of SH3 domains by comparing the binding properties of wild-type Src SH3 and Abl SH3 with those of a Src SH3 mutant containing a mutated arginine binding site (D99N) and Abl SH3 mutant constructs engineered to contain an arginine binding site (T98D and T98D/F91Y). We found that the D99N mutation diminished binding to most Src SH3-binding proteins in whole cell extracts; however, there was only a moderate reduction in binding to a small subset of Src SH3-binding proteins (including the Src substrate p68). p68 was shown to contain two Arg-containing Asp-99-dependent binding sites and one Asp-99-independent binding site which lacks an Arg. Moreover, substitution of Asp for Thr-98 in Abl SH3 changed the binding specificity of this domain and conferred the ability to recognize Arg-containing ligands. These results indicate that Asp-99 is important for Src SH3 binding specificity and that Asp-99-dependent binding interactions play a dominant role in Src SH3 recognition of cellular binding proteins, and they suggest the existence of two Src SH3 binding mechanisms, one requiring Asp-99 and the other independent of this residue.
机译：SH3结构域通过识别细胞蛋白上富含脯氨酸的序列基序来介导细胞内蛋白-蛋白相互作用。对与富含脯氨酸的肽配体复合的Src SH3域（Src SH3）的结构分析显示，该相互作用涉及三个结合位点：两个疏水相互作用（在SH3配体中的脂族脯氨酸二肽与SH3表面上高度保守的芳族残基之间）域）和一个盐桥（Src的Asp-99与Arg之间的配体中Pro-XX-Pro保守基序上游三个残基）。我们通过比较野生型Src SH3和Abl SH3与包含突变精氨酸结合位点（D99N）的Src SH3突变体和工程改造为包含Abl SH3突变体的Src SH3突变体的结合特性，研究了SH3结构域精氨酸结合位点的重要性。精氨酸结合位点（T98D和T98D / F91Y）。我们发现，D99N突变减少了与全细胞提取物中大多数Src SH3结合蛋白的结合。然而，与Src SH3结合蛋白的一小部分（包括Src底物p68）的结合仅适度降低。显示p68含有两个含Arg的Asp-99依赖性结合位点和一个缺少Arg的Asp-99非依赖性结合位点。此外，用Asp代替Abl SH3中的Thr-98改变了该结构域的结合特异性，并赋予了识别含Arg配体的能力。这些结果表明，Asp-99对Src SH3的结合特异性很重要，并且Asp-99依赖性结合相互作用在Src SH3识别细胞结合蛋白中起着主导作用，并且它们表明存在两种Src SH3结合机制，一个需要Asp-99和另一个独立于此残基。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号