An improved method for the synthesis of cellulose membrane-bound peptides with free C termini is useful for PDZ domain binding studies
An improved method for the synthesis of cellulose membrane-bound peptides with free C termini is useful for PDZ domain binding studies
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DOI：
10.1016/j.chembiol.2004.03.010
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发表时间：
2004-04-01
影响因子：
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通讯作者：
Volkmer-Engert, R
中科院分区：
文献类型：
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作者：
Boisguerin, P;Leben, R;Volkmer-Engert, R
SPOT合成允许平行合成和筛选成千上万的纤维素膜结合肽，以研究蛋白质组学背景下的蛋白质-蛋白质相互作用。C-末端残基的识别是PDZ结构域最常见的结合特征之一。不幸的是，大多数固体支持物结合的肽文库由于C-末端固定在固体支持物上而缺乏游离C-末端。为了克服这一限制，我们开发了一个强大的方法，基于我们以前的策略，用于产生肽与真实的C末端。为了验证这种改进的方法，我们筛选了6223 C末端与突触营养蛋白PDZ结构域的人肽库。此外，使用相同的文库，发现了ERBIN PDZ结构域的衍生自膜蛋白和受体的新肽配体。最后，我们确定了蛋白激酶断裂点簇区域，这是已知的细胞增殖和致癌转化的负调节因子，作为ERBIN配体。
SPOT synthesis permits parallel synthesis and screening of thousands of cellulose membrane-bound peptides to study protein-protein interactions in a proteomic context. Recognition of C-terminal residues is one of the most common binding features of PDZ domains. Unfortunately, most solid support-bound peptide libraries lack a free C terminus due to C-terminal fixation on the solid support. To overcome this restriction we developed a robust methodology based on our previous strategy for generating peptides with authentic C termini. To validate this improved method, we screened a human peptide library of 6223 C termini with the syntrophin PDZ domain. Furthermore, using the same library, new peptide ligands derived from membrane proteins and receptors were found for the ERBIN PDZ domain. Finally, we identified the protein kinase breakpoint cluster region, which is known as a negative regulator of cell proliferation and oncogenic transformation, as an ERBIN ligand.