Abstract
As biology approaches the 50th year of deciphering the DNA code, the next frontier toward understanding cell function has protein biochemistry in the form of structural and functional proteomics. To accomplish the needs of proteomics, novel strategies must be devised to examine the gene products or proteins, emerged as en masse. The authors have developed a high-throughput system for the expression and purification of eukaryotic proteins to provide the resources for structural studies and protein functional analysis. The long-term objective is to overexpress and purify thousands of proteins encoded by the human genome. This library of proteins—the human proteome—can be arrayed in addressable format in quantities and purities suitable for high-throughput studies. Critical technology involved in efficiently moving from genome to proteome includes parallel sample handling, robust expression, and rapid purification procedures. Automation of these processes is essential for the production of thousands of recombinant proteins and the reduction of human error.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Holt, L. J., Enever C., deWildt R. M. T., et al. (2000) The use of recombinant antibodies in proteomics. Curr. Opin. Biotechnol. 11, 445–449.
Zhu, H. and Synder, M. (2001) Protein arrays and microarrays. Curr. Opin. Chem. Biol. 5, 40–45.
Cahill, D. J. (2001) Protein and antibody arrays and their medical applications. J. Immunol. Methods 250, 81–91.
Reineke, U., Volkmer-Engert, R., and Schneider-Mergener, J. (2001) Applications of peptide arrays prepared by the SPOT-technology. Curr. Opin. Biotechnol. 12, 59–64.
Albala, J. S. (2001) Array-based proteomics: the latest chip challenge. Expert Rev. Mol. Diagn. 1, 145–152.
Schena, M., Shalon, D., Heller, R., et al. (1996) Parallel human genome analysis: microarray-based expression monitoring of 1000 genes. Proc. Natl. Acad. Sci. USA 93, 10,614–10,619.
Christendat, D., Yee, A., Dharamsi, A., et al. (2000) Structural proteomics of an archaeon. Nat. Struct. Biol. 7, 903–909.
Larsson, M., Graslund, S., Yuan, L., et al. (2002) High-throughput protein expression of cDNA products as a tool in functional genomics. J. Biotechnol. 80, 143–157.
Bussow, K., Cahill, D., Nietfeld, W., et al. (1998) A method for global protein expression and antibody screening on high-density filters of an arrayed cDNA library. Nucleic Acids Res. 26, 5007–5008.
Braun, P., Hu, Y., Shen, B., et al. (2002) Proteome-scale purification of human proteins from bacteria. Proc. Natl. Acad. Sci. USA 99, 2654–2659.
Lueking, A., Holz, C., Gotthold, C., et al. (2000) A system for dual protein expression in Pichia pastoris and Escherichia coli. Protein Expr. Purif. 20, 372–378.
Gilbert, M. and Albala, J. S. (2002). Accelerating code to function: sizing up the protein production-line. Curr. Opin. Chem. Biol. 6, 102–105.
Albala, J. S., Franke, K., McConnell, I. R., et al. (2000) From genes to proteins: high-throughput expression and purification of the human proteome. J. Cell. Biochem. 80, 187–191.
Lennon, G., Auffray, C., Polymeropoulos, M., et al. (1996) The I.M.A.G.E. Consortium: an integrated molecular analysis of genomes and their expression. Genomics 33, 151–152.
MacBeath, G. and Schreiber, S. L. (2000) Printing proteins as microarrays for high-throughput functional determination. Science 289, 1760–1763.
Rubinfeld, B., Munemitsu, S., Clark, R., et al. (1991) Molecular cloning of a GTPase activating protein specific for the Krev-1 protein p21rap1. Cell 65, 1033–104.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2004 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Gilbert, M., Edwards, T.C., Albala, J.S. (2004). Protein Expression Arrays for Proteomics. In: Fung, E.T. (eds) Protein Arrays. Methods in Molecular Biology, vol 264. Humana Press. https://doi.org/10.1385/1-59259-759-9:015
Download citation
DOI: https://doi.org/10.1385/1-59259-759-9:015
Publisher Name: Humana Press
Print ISBN: 978-1-58829-255-1
Online ISBN: 978-1-59259-759-8
eBook Packages: Springer Protocols