申请人:——
公开号:US20030018169A1
公开(公告)日:2003-01-23
The present invention relates to methods and compositions for lipid matrix-assisted chemical ligation and synthesis of membrane polypeptides that are incorporated in a lipid matrix. The invention is exemplified in production of a prefolded membrane polypeptide embedded within a lipid matrix via stepwise chemoselective chemical ligation of unprotected peptide segments, where at least one peptide segment is embedded in a lipid matrix. Any chemoselective reaction chemistry amenable for ligation of unprotected peptide segments can be employed. Suitable lipid matrices include liposomes, micelles, cell membrane patches and optically isotropic cubic lipidic phase matrices. Prefolded synthetic and semi-synthetic membrane polypeptides synthesized according to the methods and compositions of the invention also permit site-specific incorporation of one or more detectable moieties, such as a chromophore, which can be conveniently introduced during synthesis. The methods and compositions of the invention have multiple uses. For example, they can be used to assay ligand binding to membrane polypeptides and domains comprising a receptor, and thus are extremely useful for structure/function studies, drug screening/selection/design, and diagnostics and the like, including high-throughput applications. The methods and compositions of the invention are particularly suited for FRET analyses of previously inaccessible membrane polypeptides.
本发明涉及脂质基质辅助化学键合和合成嵌入脂质基质中的膜多肽的方法和组合物。本发明的实例是通过对未受保护的肽段进行逐步化学选择性连接,生产出嵌入脂质基质中的预折叠膜多肽,其中至少有一个肽段嵌入脂质基质中。可以采用任何适合于未受保护肽段连接的化学选择反应。合适的脂质基质包括脂质体、胶束、细胞膜片和光学各向同性立方脂相基质。根据本发明的方法和组合物合成的预折叠合成和半合成膜多肽还可在特定位点加入一个或多个可检测的分子,如发色团,这些分子可在合成过程中方便地引入。本发明的方法和组合物有多种用途。例如,它们可用于检测配体与膜多肽和包含受体的结构域的结合,因此在结构/功能研究、药物筛选/选择/设计、诊断等方面非常有用,包括高通量应用。本发明的方法和组合物尤其适用于以前无法获得的膜多肽的 FRET 分析。