Activity-Based High-Throughput Screening of Enzymes by Using a DNA Microarray

摘要

Remarkable advances in genomics have been accomplished, including the development and application of the DNA microarray technology, and the recent completion of the Human Genome Project. Consequently, the enormous amount of genetic information at an organism's transcriptional level is now becoming available. This situation has created the tremendous challenge to develop new techniques capable of studying the between 100 000 and 1 000 000 functionally expressed proteins estimated in the human proteome alone. Despite numerous innovations, to date, no single proteomic technique can encompass the diverse functionalities of proteins in a proteome. For example, two-dimensional gel electrophoresis (2D-GE), coupled with mass spectrometry, is primarily used to study the relative abundance, but not enzymatic activity, of proteins expressed in a biological sample. Other techniques have been developed for proteome-wide analysis of protein structure, localization, and interactions. One of them, the protein microarray, offers the chance to study a variety of protein activities in a large scale. However, the development of this technology is largely hampered by the cost and effort needed to generate many functional proteins in sufficient purity, as well as a lack of microarray-compatible assays available to screen for different proteins, for example, enzymes spotted in a microarray.