Proteomics cardiac

The majority of proteomic investigations employ two-dimensional gel electrophoresis (2-D) with immobilized pH gradients (IPGs) to separate the proteins in a sample and combine this with mass spectrometry (MS) technologies to identify proteins. 

Edited by M. Hamacher, K. Marcus, K. Stiihler, A. van Hall, B. Warscheid, H. E. Meyer Copyright 2006 Wiley-VCH Verlag GmbH Co. KGaA, Weinheim ISBN 3-527-31226-9 

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In this chapter we will review proteomic investigations of cardiac proteins and focus on their application to the study of heart disease in the human and in animal models of cardiac dysfunction. The majority of these studies of the cardiac proteome have involved protein separation, visualisation and quantitation using the traditional 2-DE approach combined with protein identification by mass spectrometry. These essential technologies will be briefly described. However, there is increasing interest in using alternative gel-free techniques based on mass spectrometry or protein arrays for high throughput proteomics. These alternative approaches will be introduced, but further details can be found in Chapter 2 of this volume by Michel Faupel. 

Arnott D et al. An integrated approach to proteome analysis identification of proteins associated with cardiac hypertrophy. Anal Biochem 1998 258 1-18. 

ARNOTT, D., O CONNELL, K.L., KING, K.L., STULTS, J.T., An integrated approach to proteome analysis Identification of proteins associated with cardiac hypertrophy, Anal. Biochem., 1998,258, 1-18. 

There are four gel protein databases of cardiac proteins, established by three independent groups, that can be accessed via the World Wide Web (Table 16.1). These databases facilitate proteomic research into heart diseases containing information on several hundred cardiac proteins that have been identified by protein chemical methods. They all conform to the rules for federated 2-D protein databases (Appel et al., 1996). In addition, 2-D protein databases for other mammals, such as the mouse, rat (li et al., 1999), dog (Dimn et al., 1997), pig and cow, are also under construction to support work on animal models of heart disease and heart failure. 

To date, proteomic investigations into human heart disease have centered on dilated cardiomyopathy (DCM). DCM is a disease of unknown etiology, characterized by impaired systolic function resulting in heart failure. Known contributory factors of DCM are viral infections, cardiac-specific autoantibodies, toxic agents, genetic factors, and sustained alcohol abuse. As many as 100 cardiac proteins... 

Mitochondria are involved in a variety of cellular processes. Their primary role is the production of ATP, but they are also involved in ionic homeostasis, apoptosis, oxidation of carbohydrates and fatty acids, and a variety of other catabolic and anabolic pathways. Mitochondrial dysfunction, therefore, can cause a variety of diseases including heart disease (Hirano et al., 2001). The characterization of the mitochondrial proteome could provide interesting new insights into cardiac dysfimction in heart disease (Lopez and Melov, 2002). 

Cell culture systems are ideal for detailed proteomic investigations of responses in protein expression to controlled stimuli. This is because they should provide defined systems with much lower inherent variability between samples, particularly if established cell lines are used. However, cells that are maintained in culture respond by alterations in their gene pattern, and consequently the protein expression, such that it can be quite different from that found in vivo. This process can occur quite rapidly in primary cultures of cells established from tissue samples and is even more profound in cells maintained long-term, particularly where transformation has been used to establish immortal cell lines. Cardiac myocytes can pose an even bigger challenge. While neonatal cardiac myocytes can be maintained and grown in vitro, adult cells are terminally differentiated and can be maintained for relatively short times in vitro but are not capable of cell division. 

Arnott et al. (1998) have used phenylephrine treated neonatal rat cardiac myocytes as a model of cardiac hypertrophy for proteomic analysis. In this 2-D-based study, 11 protein spots displayed statistically significant changes in expression upon induction of hypertrophy. Of these, eight showed higher expression and three were decreased in abundance in hypertrophied cells. All of... 

Proteomic Characterization of Cardiac Antigens in Heart Disease and Transplantation... 

Petricoin EF, Rajapaske V, Herman EH, Arekani AM, Ross S, Johann D, Knapton A, Zhang J, Hitt BA, Conrads TP, Veenstra TD, Liotta LA, Sistare FD. Toxicoproteomics Seram proteomic pattern diagnostics for early detection of drag induced cardiac toxicities and cardioprotection. Toxicol Pathol 2004 32 (Suppl l) 122-30. 

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