MacBeath G, Schreiber S L
Center for Genomics Research, Harvard University, 16 Divinity Avenue, Cambridge, MA 02138, USA.
Science. 2000 Sep 8;289(5485):1760-3. doi: 10.1126/science.289.5485.1760.
Systematic efforts are currently under way to construct defined sets of cloned genes for high-throughput expression and purification of recombinant proteins. To facilitate subsequent studies of protein function, we have developed miniaturized assays that accommodate extremely low sample volumes and enable the rapid, simultaneous processing of thousands of proteins. A high-precision robot designed to manufacture complementary DNA microarrays was used to spot proteins onto chemically derivatized glass slides at extremely high spatial densities. The proteins attached covalently to the slide surface yet retained their ability to interact specifically with other proteins, or with small molecules, in solution. Three applications for protein microarrays were demonstrated: screening for protein-protein interactions, identifying the substrates of protein kinases, and identifying the protein targets of small molecules.
目前正在进行系统性的努力，以构建用于重组蛋白高通量表达和纯化的克隆基因定义集。为便于后续对蛋白质功能的研究，我们开发了微型化检测方法，该方法可容纳极低的样本量，并能快速、同时处理数千种蛋白质。一台设计用于制造互补DNA微阵列的高精度机器人被用于以极高的空间密度将蛋白质点样到化学衍生的载玻片上。共价附着在载玻片表面的蛋白质仍保留其在溶液中与其他蛋白质或小分子特异性相互作用的能力。展示了蛋白质微阵列的三种应用：筛选蛋白质-蛋白质相互作用、鉴定蛋白激酶的底物以及鉴定小分子的蛋白质靶点。
