Abstract
The isolation of a given protein, free of all other biomolecules, is the primary objective of any protein purification scheme. Classical chromatographic procedures have been designed to exploit particular distinguishing features of individual target proteins, such as size, physico-chemical properties and binding affinity. Advances in molecular biology and bioinformatics have positively contributed at every level to the challenge of purifying individual proteins and more recently have led to the development of high-throughput proteomic platforms. Here, a summation of developments in the field of protein chromatography is given, coupled with a compilation of general resources and tools that are available to assist with protein purification processes.
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References
Sun, P.D., Foster, C.E., and Boyington, J.C. (2004) Overview of protein structural and functional folds. Curr. Protoc. Protein Sci. Chapter 17: Unit 17.1.
Kang, T.S. and Kini, R.M. (2009) Structural determinants of protein folding. Cell. Mol. Life Sci. 66, 2341–2361.
Bartlett, A.I. and Radford, S.E. (2009) An expanding arsenal of experimental methods yields an explosion of insights into protein folding mechanisms. Nat. Struct. Mol. Biol. 16, 582–588.
Strandberg, B. (2009) Building the ground for the first two protein structures: myoglobin and haemoglobin. J. Mol. Biol. 392, 2–10.
Cohen, E.J., Strong, L.E., Hughes, W.L., Mulford, D.J., Ashworth, J.N., Melin, M., and Taylor, H.L. (1946) Preparation and properties of serum and plasma proteins. IV. A system for the separation into fractions of the proteins and lipoprotein components of biological tissues and fluids. J. Am. Chem. Soc. 68, 459–475.
Katzen, F., Peterson, T.C., and Kudlicki, W. (2009) Membrane protein expression: no cells required. Trends Biotechnol. 27, 455–460.
Gasteiger, E., Gattiker, A., Hoogland, C., Ivanyi, I., Appel, R.D., and Bairoch A. (2003) ExPASy: the proteomics server for in-depth protein knowledge and analysis. Nucleic Acids Res. 31, 3784–3788.
Craig, R., Cortens, J.P., and Beavis, R.C. (2004) Open source system for analyzing, validating, and storing protein identification data. J. Proteome Res. 3(6), 1234–1242.
Rice, P., Longden, I., and Bleasby, A. (2000) EMBOSS: the European Molecular Biology Open Software Suite. Trends Genet. 16(6), 276–277.
Fuhrmann, M., Hausherr, A., Ferbitz, L., Schödl, T., Heitzer, M., and Hegemann, P. (2004) Monitoring dynamic expression of nuclear genes in Chlamydomonas reinhardtii by using a synthetic luciferase reporter gene. Plant Mol. Biol. 55(6), 869–881.
Stothard, P. (2000) The Sequence Manipulation Suite: JavaScript programs for analyzing and formatting protein and DNA sequences. Biotechniques 28, 1102–1104.
Kessner, D., Chambers, M., Burke, R., Agus, D., and Mallick, P. (2008) ProteoWizard: Open source software for rapid proteomics tools development. Bioinformatics 24(21), 2534–2536.
Bendtsen, J.D., Nielsen, H., von Heijne, G., and Brunak, S. (2004) Improved prediction of signal peptides: SignalP 3.0. Mol. Biol. 340, 783–795.
Blom, N., Sicheritz-Ponten, T., Gupta, R., Gammeltoft, S., and Brunak, S. (2004) Prediction of post-translational glycosylation and phosphorylation of proteins from the amino acid sequence. Proteomics 4(6), 1633–1649.
Gardy, J.L., Laird, M.R., Chen, F., Rey, S., Walsh, C.J., Ester, M., and Brinkman, F.S.L. (2005) PSORTb v.2.0: Expanded prediction of bacterial protein subcellular localization and insights gained from comparative proteome analysis. Bioinformatics 21(5), 617–623.
Guda, C. (2006) pTARGET: a web server for predicting protein subcellular localization. Nucleic Acids Res. 34 (web server issue), W210–W213.
Cserzo, M., Wallin, E., Simon, I., von Heijne, G., and Elofsson, A. (1997) Prediction of transmembrane alpha-helices in procariotic membrane proteins: the Dense Alignment Surface method. Prot. Eng. 10(6), 673–676.
Hofmann, K. and Stoffel, W. (1993) TMbase – A database of membrane spanning proteins segments. Biol. Chem. Hoppe Seyler 374, 166.
Cole, C., Barber, J.D., and Barton, G.J. (2008) The Jpred 3 secondary structure prediction server. Nucleic Acids Res. 36 (web server issue), W197–W201.
Diemand, A.V. and Scheib, H. (2004) MolTalk – a programming library for protein structures and structure analysis. BMC Bioinformatics, 5, 39.
Sippl, M.J. and Wiederstein, M. (2008) A note on difficult structure alignment problems. Bioinformatics 24, 426–427.
Kiefer, F., Arnold, K., Künzli, M., Bordoli, L., and Schwede, T. (2009). The SWISS-MODEL Repository and associated resources. Nucleic Acids Res. 37, D387–D392.
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Walls, D., McGrath, R., Loughran, S.T. (2011). A Digest of Protein Purification. In: Walls, D., Loughran, S. (eds) Protein Chromatography. Methods in Molecular Biology, vol 681. Humana Press. https://doi.org/10.1007/978-1-60761-913-0_1
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DOI: https://doi.org/10.1007/978-1-60761-913-0_1
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