Electrophoresis map

FIGURE 1.1 A 2D-gel electrophoresis map of colorectal epithelia cells proteins from the SWISS- 2DPage database (entry CATD HUMAN, primary access number P07339) accessible from http //www.expasy.org/swiss-2dpage. 

Pietrogrande, M.C., Marchetti, N., Dondi, F., Righetti, P.G. (2003). Spot overlapping in two-dimensional polyacrylamide gel electrophoresis maps relevance to proteomics. Electrophoresis 24, 217. 

D-polyacrylamide gel electrophoresis) maps of protein mixtures is discussed. 2D PAGE is considered the classical and principal tool for protein separation—prior to mass spectrometry—to achieve the main goal of proteomics, that is, a comprehensive identification and quantification of every protein present in a complex biological sample that would allow analysis of an entire intact proteome (Wilkins et al., 1997 Righetti et al., 2001 Hamdan and Righetti, 2005). 

Seow TK et al. Two-dimensional electrophoresis map of the human hepatocellular carcinoma cell line, HCC-M, and identification of the separated proteins by... 

Sanchez-Campillo, M., Bini, L., Comanducci, M., Raggiaschi, R., Marzocchi, B., Pallini, V., et al. (1999) Identification of immunoreactive proteins of Chlamydia trachomatis by Western blot analysis of a two-dimensional electrophoresis map with patient sera. Electrophoresis 20,2269-2279. 

Western blot analysis of a two-dimensional electrophoresis map with patient sera. Electrophoresis 20 2269-2279, 1999. 

Talamo F, D Ambrosio C, Arena S, Del Vecchio P, Ledda L, Zehender G, Ferrara L, Scaloni A. Proteins from bovine tissues and biological fluids Defining a reference electrophoresis map for liver, kidney, muscle, plasma and red blood cells. Proteomics 2003 3(4) 440-60. 

Haynes, R, I. Miller, R. Aebersold, M. Gemeiner, I. Eberini, M. R. Lovati, C. Manzoni, M. Vignati, and E. Gianazza. 1998. Proteins of rat serum I. Establishing a reference two-dimensional electrophoresis map by immunodetection and microbore high-performance liquid chromatography-electrospray mass spectrometry. Electrophoresis 19 1484-1492. 

An important factor in the progress of bioinformatics has been the constant increase in computer speed and memory capacity of desktop computers and the increasing sophistication of data processing techniques. The computation power of common personal computers has increased within 12 years approximately 100-fold in processor speed, 250-fold in RAM memory space and 500-fold or more in hard disk space, while the price has nearly halved. This enables acquisition, transformation, visuahsation and interpretation of large amounts of data at a fraction of the cost compared to 12 years ago. Presently, bioanalytical databases are also growing quickly in size and many databases are directly accessible via the Internet One of the first chemical databases to be placed on the Internet was the Brookha-ven protein data bank, which contains very valuable three-dimensional structural data of proteins. The primary resource for proteomics is the ExPASy (Expert Protein Analysis System) database, which is dedicated to the analysis of protein sequences and structures and contains a rapidly growing index of 2D-gel electrophoresis maps. Some primary biomolecular database resources compiled from spectroscopic data are given in Tab. 14.1. 

M. Stromqvist, Peptide mapping using combinations of size-exclusion chromatography, reversed-phase chromatography and capillary electrophoresis , 7. Chromatogr. 667 304-310(1994). 

Microtubule-associated proteins bind to microtubules in vivo and subserve a number of functions including the promotion of microtubule assembly and bundling, chemomechanical force generation, and the attachment of microtubules to transport vesicles and organelles (Olmsted, 1986). Tubulin purified from brain tissue by repeated polymerization-depolymerization contains up to 20% MAPs. The latter can be dissociated from tubulin by ion-exchange chromatography. The MAPs from brain can be resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). 

The extraordinary complexity of human genes and their products has encouraged the development of extremely high-resolution analytical methods.75 Capillary electrophoresis is competitive with slab gel methods, with resolution up to the order of about 1,000 base pairs for sequencing, sizing, and detection of mutation. Reversed phase HPLC is useful for restriction digest mapping and MALDI-MS up to about 1000 base pairs. 

Palmieri, R., Peptide mapping by capillary electrophoresis, Appl. Brief DS-774, Beckman Instruments, Fullerton, CA, 1990. 

Suzuki, S., Kakehi, K., and Honda, S., Two-dimensional mapping of N-gly-cosidically linked asialo-oligosaccharides from glycoproteins as reductively pyridylaminated derivatives using dual separation modes of high-performance capillary electrophoresis, Anal. Biochem., 205, 227, 1992. 

Bonneil, E., Mercier, M. and Waldron, K.C., Reproducibility of a solid-phase trypsin microreactor for peptide mapping by capillary electrophoresis, Anal. Chim. Acta, 404, 29, 2000. 

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