SBIR PHASE I: Metal Cluster and Combined Flourescent/Gold Labels for Peptide Synthesis

在这个小企业创新研究的第一阶段项目中，十一金、1.4 nm的“纳米金”团簇配合物、结合荧光和金团簇标签以及直径1.8 - 3.6 nm的铂团簇配合物将被共价偶联到适合于支撑肽合成的保护氨基酸上。将制备l -半胱氨酸、l -赖氨酸和甘氨酸的类似物，以及功能化的标记试剂，将簇标记附加到n端和c端，并在自动肽合成过程中用于在合成肽的编程位置插入标记。天然氨基酸的手性将被保留，以保持最大的肽生物活性。这些标签将通过配位体的系统变化，在合成和脱保护条件下获得最大的溶解度和稳定性。制备组蛋白H1蛋白、HMG-D和HMG-Z蛋白c端标记的肽类似物，并将其微注射到活细胞中。荧光共聚焦和电子显微镜将研究它们在细胞核内的分布、功能和大分子定位。标签也将被纳入合成的激素肽促白素释放激素（LHRH）类似物中，然后在膜印迹和微滴板检测中研究其作为免疫测定探针的敏感性和特异性。该试剂将被电子显微镜、细胞和分子生物学家广泛用于制备具有最高分辨率、保留生物活性和灵敏度的探针。同时制备标记和未标记肽的策略将减少探针制备时间和成本。快速增长的应用存在于选择性标记组合肽。这些试剂也可用于制备新型杂化材料，其中金属纳米颗粒的独特电子和催化特性被编程为特定位点的新生物材料，以产生新的大分子器件。

9561345 Powell In this Small Business Innovation Research Phase I project, Undecagold, 1.4 nm "Nanogold" cluster complexes, combined fluorescent and gold cluster labels, and platinum cluster complexes 1.8 to 3.6 nm in diameter will be covalently conjugated to protected amino acids suitable for supported peptide synthesis. Analogs of L-cysteine, L-lysine and glycine will be prepared, in addition to labeling reagents functionalized to append the cluster labels to the N-terminal and C-terminal, and will be used to insert the labels at programmed positions in synthetic peptides during automated peptide synthesis. Chirality of the natural amino acids will be preserved to retain maximum peptide bioactivity. The labels will be synthetically modified for maximum solubility and stability towards synthesis and deprotection conditions by systematic variations of their coordinated ligands. Labeled peptide analogs of the C-terminal region of histone H1 protein, HMG-D and HMG-Z proteins will be prepared and microinjected into living cells. Their distribution, function and macromolecular localization within the cell nucleus will be studied by fluorescence confocal and electron microscopy. Labels will also be incorporated into synthetic analogs of the hormone peptide leuteinizing hormone releasing hormone (LHRH), then their sensitivity and specificity as immunoassay probes will be studied in membrane blot and microtiter plate tests. The reagents will be widely used by electron microscopists, cell and molecular biologists to prepare probes with the highest possible resolution, retention of bioactivity and sensitivity. The strategy for preparing labeled and unlabeled peptides simultaneously will reduce probe preparation time and cost. Rapidly growing applications exist in the selective labeling of combinatorial peptides. These reagents may also be used to prepare novel hybrid materials in which the unique electronic and catalytic properties of metal nanoparticles are programmed site-specif ically into new biomaterials to yield new macromolecular devices.