Printing proteins as microarrays for high-throughput function determination.
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.
MacBeath
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Center for Genomics Research, Harvard University, 16 Divinity Avenue, Cambridge, MA 02138, USA. gavin_macbeath@harvard.edu
Schreiber
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SL
D001671:Biochemistry / Q000379:methods*; D001710:Biotin / Q000378:metabolism; D004076:Digoxigenin / Q000378:metabolism; D005453:Fluorescence; D005456:Fluorescent Dyes; D008024:Ligands; D015336:Molecular Probe Techniques; D010766:Phosphorylation; D010879:Piperazines / Q000494:pharmacology; D011485:Protein Binding; D017510:Protein Folding; D011494:Protein Kinases / Q000378:metabolism*; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D012371:Robotics; D012710:Serum Albumin, Bovine
C413289:AF 1497; D005456:Fluorescent Dyes; D008024:Ligands; D010879:Piperazines; D011506:Proteins; D012710:Serum Albumin, Bovine; D001710:Biotin; D011494:Protein Kinases; D004076:Digoxigenin