Van Eyk, Dunn - Proteomic and Genomic Analysis of Cardiovascular Disease - 2003 (522919), страница 54
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As in bovine DCM, expression of the enzyme UCH was more than 8-fold elevated at the protein level andmore than 5-fold elevated at the mRNA level in human DCM. Moreover, this increased expression of UCH was shown by immunocytochemistry to be associated11.8 Summarywith the myocytes which do not exhibit detectable staining in control hearts. Overall protein ubiquitination was increased 5-fold in DCM relative to control heartsand using a selective affinity purification method we were able to demonstrate enhanced ubiquitination of a number of distinct proteins in DCM hearts. We haveidentified a number of these proteins by mass spectrometry.
Interestingly many ofthese proteins were the same proteins that we have previously found to be present at reduced abundance in DCM hearts [53]. This new evidence strengthensour hypothesis that inappropriate ubiquitin conjugation leads to proteolysis anddepletion of certain proteins in the DCM heart and may contribute to loss of normal cellular function in the diseased heart.11.7.4Proteomic Characterization of Cardiac Antigens in Heart Disease and TransplantationProteomics can be utilized to identify cardiac-specific antigens that elicit antibodyresponses in heart disease and following cardiac transplantation. This approachmakes use of Western blot transfers of 2DE separations of cardiac proteins. Theseare probed with patient serum samples and developed using appropriately conjugated anti-human immunoglobulins.
This strategy has revealed several cardiacantigens that are reactive with autoantibodies in DCM [70, 71] and myocarditis[72]. Cardiac antigens that are associated with antibody responses following cardiac transplantation have also been characterized and may be involved in acute [73]and chronic [74] rejection.11.8SummaryThe use of proteomics in cardiovascular research has enabled the identificationand characterization of differential protein expression in heart disease. By complementing genomic-based approaches, new insights into complex cellular processeswill improve our understanding of cardiac dysfunction. Proteomics should allowan understanding of disease at the molecular level and provide the means foridentifying novel diagnostic disease markers.
This in turn will accelerate the development of new therapeutic approaches.18919011 Proteomics, a Step Beyond Genomics11.9References1234567891011O’Farrell, P. H. High resolution two-dimensional electrophoresis of proteins. J.Biol. Chem. 1975, 250, 4007–4021.Anderson, N.
G., Anderson, L. The Human Protein Index. Clin.Chem. 1982, 28,739–748.Anderson, N. G., Matheson, A., Anderson, N. L. Back to the future: the humanprotein index (HPI) and the agenda forpost-proteomic biology. Proteomics. 2001,1, 3–12.Wasinger, V. C., Cordwell, S. J., CerpaPoljak, A., Yan, J. X., et al. Progresswith gene-product mapping of the Mollicutes: Mycoplasma genitalium. Electrophoresis. 1995, 16, 1090–1094.Wilkins, M.
R., Sanchez, J. C., Gooley,A. A., Appel, R. D., et al. Progress withproteome projects: why all proteins expressed by a genome should be identified and how to do it. Biotechnol. Genet.Eng Rev. 1996, 13, 19–50.Anderson, L., Seilhamer, J. A comparison of selected mRNA and protein abundances in human liver. Electrophoresis.1997, 18, 533–537.Haynes, P. A., Gygi, S. P., Figeys, D., Aebersold, R. Proteome analysis: biologicalassay or data archive? Electrophoresis1998, 19, 1862–1871.Dunn, M. J. Two-dimensional polyacrylamide gel electrophoresis.
Adv. Electrophoresis. 1987, 1, 1–109.Rabilloud, T., Gianazza, E., Catto, N.,Righetti, P. G. Amidosulfobetaines, afamily of detergents with improved solubilization properties: application for isoelectric focusing under denaturing conditions. Anal. Biochem. 1990, 185, 94–102.Rabilloud, T. Use of thiourea to increase the solubility of membrane proteins in two- dimensional electrophoresis. Electrophoresis. 1998, 19, 758-760.Bjellqvist, B., Ek, K., Righetti, P.
G.,Gianazza, E., et al. Isoelectric focusingin immobilized pH gradients: principle,methodology and some applications. J.Biochem. Biophys. Methods. 1982, 6, 317–339.1213141516171819Gorg, A., Boguth, G., Obermaier, C.,Posch, A., et al. Two-dimensional polyacrylamide gel electrophoresis with immobilized pH gradients in the first dimension (IPG-Dalt): the state of the art andthe controversy of vertical versus horizontal systems. Electrophoresis. 1995, 16,1079–1086.Corbett, J. M., Dunn, M.
J., Posch, A.,Gorg, A. Positional reproducibility ofprotein spots in two-dimensional polyacrylamide gel electrophoresis using immobilised pH gradient isoelectric focusing in the first dimension: an interlaboratory comparison. Electrophoresis.1994, 15, 1205–1211.Blomberg, A., Blomberg, L., Norbeck,J., Fey, S. J., et al. Interlaboratory reproducibility of yeast protein patterns analyzed by immobilized pH gradient twodimensional gel electrophoresis. Electrophoresis. 1995, 16, 1935–1945.Craven, R.
A., Jackson, D. H., Selby,P. J., Banks, R. E. Increased protein entrytogether with improved focussing usinga combined IPGphor/Multiphorapproach. Proteomics. 2002, 2, 1061–1063.Gorg, A., Obermaier, C., Boguth, G.,Harder, A., et al. The current state oftwo-dimensional electrophoresis with immobilized pH gradients. Electrophoresis.2000, 21, 1037–1053.Choe, L. H., Lee, K. H. A comparison ofthree commercially available isoelectricfocusing units for proteome analysis: themultiphor, the IPGphor and the proteanIEF cell.
Electrophoresis. 2000, 21, 993–1000.Klose, J., Kobalz, U. Two-dimensionalelectrophoresis of proteins: an updatedprotocol and implications for a functionalanalysis of the genome. Electrophoresis.1995, 16, 1034–1059.Westbrook, J. A., Yan, J. X., Wait, R.,Welson, S. Y., et al.
Zooming-in on theproteome: very narrow-range immobilised pH gradients reveal more proteinspecies and isoforms. Electrophoresis.2001, 22, 2865–2871.11.9 References202122232425262728Wildgruber, R., Harder, A., Obermaier, C., Boguth, G., et al. Towardshigher resolution: two-dimensional electrophoresis of Saccharomyces cerevisiaeproteins using overlapping narrow immobilized pH gradients. Electrophoresis.2000, 21, 2610–2616.Wasinger, V. C., Bjellqvist, B., Humphery-Smith, I. Proteomic ‘contigs’ ofOchrobactrum anthropi, application ofextensive pH gradients.
Electrophoresis.1997, 18, 1373–1383.Gorg, A., Obermaier, C., Boguth, G.,Csordas, A., et al. Very alkaline immobilized pH gradients for two-dimensionalelectrophoresis of ribosomal and nuclearproteins. Electrophoresis. 1997, 18, 328–337.Cordwell, S. J., Nouwens, A. S., Verrills, N. M., Basseal, D. J., et al.
Subproteomics based upon protein cellular location and relative solubilities in conjunction with composite two-dimensionalelectrophoresis gels. Electrophoresis. 2000,21, 1094–1103.Neuhoff, V., Arold, N., Taube, D., Ehrhardt, W. Improved staining of proteinsin polyacrylamide gels including isoelectric focusing gels with clear backgroundat nanogram sensitivity using CoomassieBrilliant Blue G-250 and R-250. Electrophoresis. 1988, 9, 255–262.Switzer, R. C., III, Merril, C.
R., Shifrin, S. A highly sensitive silver stain fordetecting proteins and peptides in polyacrylamide gels. Anal. Biochem. 1979, 98,231–237.Shevchenko, A., Wilm, M., Vorm, O.,Mann, M. Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal. Chem. 1996, 68, 850–858.Yan, J. X., Wait, R., Berkelman, T., Harry, R., et al. A modified silver stainingprotocol for visualization of proteinscompatible with matrix-assisted laser desorption/ionization and electrospray ionization-mass spectrometry.
Electrophoresis.2002, 21, 3666–3672.Sinha, P., Poland, J., Schnolzer, M., Rabilloud, T. A new silver staining apparatus and procedure for matrix-assisted laser desorption/ionization-time of flight293031323334353637analysis of proteins after two-dimensional electrophoresis.
Proteomics. 2001,1, 835–840.Patton, W. F. A thousand points of light:the application of fluorescence detectiontechnologies to two-dimensional gel electrophoresis and proteomics. Electrophoresis. 2000, 21, 1123–1144.Lopez, M. F., Berggren, K., Chernokalskaya, E., Lazarev, A., et al.
A comparison of silver stain and SYPRO RubyProtein Gel Stain with respect to proteindetection in two-dimensional gels andidentification by peptide mass profiling.Electrophoresis. 2000, 21, 3673–3683.Berggren, K. N., Schulenberg, B., Lopez, M. F., Steinberg, T. H., et al. An improved formulation of SYPRO Ruby protein gel stain: Comparison with the original formulation and with a ruthenium IItris (bathophenanthroline disulfonate)formulation.
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