Printing proteins as microarrays for high-throughput function determination.

By G. MacBeath, Stefan Schreiber. Science, 2000

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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.

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Citations in the corpus

Subcellular localization of the yeast proteome. (2002, background): “…et al. 2000; Ito et al. 2001; Tong et al. 2002), immunoprecipitation/mass spectrometric analysis of protein complexes (Gavin et al. 2002; Ho et al. 2002), and protein microarray technologies (MacBeath and Schreiber 2000; Zhu et al. 2000, 2001) are yielding unprecedented quantities of protein data.”
Deciphering Protein–Protein Interactions. Part I. Experimental Techniques and Databases (2007, methodology): “Protein microarrays [114–116] þ In vitro Physical interaction (complex) Identification”
Fabrication of silicon nanowire devices for ultrasensitive, label-free, real-time detection of biological and chemical species (2006, background): “The reactive aldehyde groups on the SiNW surfaces can be used to couple proteins (through lysine residues”
Designing materials for biology and medicine (2004, background): “Alternative approaches to the fabrication of protein microarrays have used aldehydic surfaces to capture protein-bound lysine side chain”
Deep mutational scanning: assessing protein function on a massive scale (2011, methodology): “Higher-throughput methods such as protein arrays are useful, but at present are limited to hundreds or a few thousand proteins [8–10].”
Discrete logic modelling as a means to link protein signalling networks with functional analysis of mammalian signal transduction (2009, methodology): “PINs are usually created using data from bibliome mining (Chatr-Aryamontri et al, 2007; Kerrien et al, 2007), large-scale affinity purification (Köcher and Superti-Furga, 2007), protein arrays (MacBeath and Schreiber, 2000), and two-hybrid screening (Rual et al, 2005) or genetic interactions (Jansen”
Harnessing naturally randomized transcription to infer regulatory relationships among genes (2007, background): “Background It is now possible to measure DNA variation, RNA expression levels, and protein expression levels from thousands of genes in a given biologic sample [1-3].”
Molecular Biologist's Guide to Proteomics (2002, methodology): “Proteins were immobilized by being covalently attached to glass microscope slides, and the protein microarrays were shown to be capable of interacting with other proteins, small molecules, and enzyme substrates (98).”
A quantitative protein interaction network for the ErbB receptors using protein microarrays (2006, background): “While it is a daunting task to study the interaction of 159 proteins with 66 peptides, protein microarray technology facilitates such large-scale analyses 7,8 .”