用于氨基酸分析和蛋白质测序的微阵列
Microarrays for Amino Acid Analysis and Protein Sequencing
摘要：
本发明描述了一种方法，其中氨酰-tRNA合成酶系统用于产生非常小的装置，用于定量分析蛋白质中存在的氨基酸。该方法的基础是20种合成酶和/或对不同氨基酸特异的tRNA中的每一种在空间上分离（例如在微小的腔室中或在表面上）。以空间分辨的方式监测由所有20种合成酶催化的反应。每个单独定位的合成酶或tRNA将发出其同源氨基酸的信号。合成酶反应可以使用连续光谱测定法来监测。或者，由于延伸因子Tu;GTP（EF-Tu;GTP）特异性结合所有AA-tRNA，因此可以使用配体测定来监测由合成酶催化的氨酰化反应。建议用于氨基酸分析的微阵列。另外，氨基酸分析阵列可以与氨肽酶或羧肽酶二聚体整合以产生能够以飞摩尔-阿摩尔水平产生N-或C-末端序列数据的小型化酶促测序仪是可能的。许多样品在一个自动化的方式并行处理的可能性进行了讨论。
英文摘要：
A method is described where the aminoacyl-tRNA synthetase system is used to create very small devices for quantitative analysis of the amino acids that occur in proteins. The basis of the method is that each of the 20 synthetases and/or a tRNA specific for a different amino acid is separated spatially (e.g. in tiny chambers or on a surface). The reactions catalyzed by all 20 synthetases are monitored in a spatially resolved manner. Each separately positioned synthetase or tRNA will signal its cognate amino acid. The synthetase reactions can be monitored using continuous spectroscopic assays. Alternatively, since elongation factor Tu;GTP (EF-Tu;GTP) specifically binds all AA-tRNAs, the aminoacylation reactions catalyzed by the synthetases can be monitored using ligand assays. Microarrays for amino acid analysis are suggested. Additionally, it is possible that amino acid analysis arrays can be integrated with aminopeptidase or carboxypeptidase digestions to produce miniaturized enzymatic sequenators capable of generating either N- or C-terminal sequence data at femtomole-attomole levels. The possibility of parallel processing of many samples in an automated manner is discussed.
机构、总结、中文摘要均由AI分析而来，可能存在错误，请以论文正文为准。
