Bovine dilated cardiomyopathy: Proteomic analysis of an animal model of human dilated cardiomyopathy

Bovine hereditary dilated cardiomyopathy (bCMP) is endemic in Switzerland and hearts from diseased animals display important clinical and biochemical similarities to human DCM. Recent research has identified at least one protein (myoglobin) to be significantly reduced in bovine DCM. Using a proteomic approach, we have separated over 1125 protein species from bovine ventricular tissue. Gel analysis and protein characterisation have identified a number of proteins whose abundance is significantly altered in bovine DCM. Twenty‐four proteins are of decreased abundance in diseased tissue, whilst 11 proteins are of increased abundance in the diseased state. A combination of amino acid compositional analysis, peptide mass profiling, N‐terminal microsequencing and MultiIdent (http://www.expasy.ch/sprot/multiident.html) has been employed in order to elucidate the identities of the differentially expressed proteins. Using these techniques we have currently determined the identity of 12 of the 35 altered proteins. We have also detected three proteins that are differentially expressed in genotypically diseased but phenotypically normal animals, identifying a possible mechanism for the onset of the disease. The possibility that inappropriate ubiquination of proteins plays an important role in the disease is discussed. A database of bovine proteins is currently being established. The identity of the proteins affected, together with a comparison of the human and bovine expression patterns, is displayed.

[1]  F. Müller,et al.  Bovine hereditary cardiomyopathy: an animal model of human dilated cardiomyopathy. , 1995, Journal of molecular and cellular cardiology.

[2]  C. Lobsiger,et al.  [Pathology of bovine cardiomyopathy]. , 1990, Schweizer Archiv fur Tierheilkunde.

[3]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[4]  J. Weil,et al.  Reduction of myocardial myoglobin in bovine dilated cardiomyopathy. , 1997, Journal of molecular and cellular cardiology.

[5]  A. Clark,et al.  The role of proteases in experimental glucocorticoid myopathy , 1981, Muscle & nerve.

[6]  M. Dunn,et al.  Protein changes observed in pacing‐induced heart failure using two‐dimensional electrophoresis , 1998, Electrophoresis.

[7]  N. Alpert,et al.  Alterations in sarcoplasmic reticulum gene expression in human heart failure. A possible mechanism for alterations in systolic and diastolic properties of the failing myocardium. , 1993, Circulation research.

[8]  S. Schiaffino,et al.  Myosin Isoenzymes in Normal and Hypertrophied Human Ventricular Myocardium , 1983, Circulation research.

[9]  R D Appel,et al.  Melanie II – a third‐generation software package for analysis of two‐dimensional electrophoresis images: I. Features and user interface , 1997, Electrophoresis.

[10]  A. Goldberg.  Functions of the proteasome: the lysis at the end of the tunnel. , 1995, Science.

[11]  R D Appel,et al.  Melanie II – a third‐generation software package for analysis of two‐dimensional electrophoresis images: II. Algorithms , 1997, Electrophoresis.

[12]  J. Clark,et al.  Inappropriate ubiquitin conjugation: a proposed mechanism contributing to heart failure. , 1997, Cardiovascular research.

[13]  M. Bristow,et al.  Calcium antagonist binding sites in failing and nonfailing human ventricular myocardium. , 1990, Biochemical pharmacology.

[14]  P. Tschudi,et al.  Herz‐ und kreislaufphysiologische Untersuchungen an Rindern mit und ohne Kardiomyopathie , 1989 .

[15]  S. Coughlin,et al.  Idiopathic dilated cardiomyopathy. , 1994, The New England journal of medicine.

[16]  M J Dunn,et al.  The human myocardial two‐dimensional gel protein database: Update 1994 , 1994, Electrophoresis.

[17]  D. Hochstrasser,et al.  Improved and simplified in‐gel sample application using reswelling of dry immobilized pH gradients , 1997, Electrophoresis.

[18]  M. Yacoub,et al.  Cardiac protein abnormalities in dilated cardiomyopathy detected by two‐dimensional polyacrylamide gel electrophoresis , 1998, Electrophoresis.

[19]  S. Hanash,et al.  Elimination of point streaking on silver stained two‐dimensional gels by addition of iodoacetamide to the equilibration buffer , 1987 .

[20]  T K Attwood,et al.  OWL--a non-redundant composite protein sequence database. , 1994, Nucleic acids research.

[21]  D. Hochstrasser,et al.  From Proteins to Proteomes: Large Scale Protein Identification by Two-Dimensional Electrophoresis and Arnino Acid Analysis , 1996, Bio/Technology.

[22]  J. Gwathmey,et al.  Experimental aspects of cardiomyopathy , 1993 .

[23]  M R Wilkins,et al.  Characterisation of proteins from two‐dimensional electrophoresis gels by matrix‐assisted laser desorption mass spectrometry and amino acid compositional analysis , 1996, Electrophoresis.

[24]  A. Ciechanover,et al.  The ubiquitin‐mediated proteolytic pathway: mechanisms of recognition of the proteolytic substrate and involvement in the degradation of native cellular proteins , 1994, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[25]  K. Boheler,et al.  Altered sarcoplasmic reticulum Ca2(+)-ATPase gene expression in the human ventricle during end-stage heart failure. , 1990, The Journal of clinical investigation.

[26]  A. Katz,et al.  The myocardium in congestive heart failure. , 1989, The American journal of cardiology.

[27]  K. Jakobs,et al.  Increase of Gi alpha in human hearts with dilated but not ischemic cardiomyopathy. , 1990, Circulation.

[28]  V. Neuhoff,et al.  Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G‐250 and R‐250 , 1988, Electrophoresis.

[29]  G. Sávay,et al.  Histochemical evidence for the role of Ca2+ and neutral protease in the development of the subacute myopathy induced by organophosphorous compounds. , 1983, Acta histochemica.

[30]  Thierry Rabilloud,et al.  Sample application by in‐gel rehydration improves the resolution of two‐dimensional electrophoresis with immobilized pH gradients in the first dimension , 1994, Electrophoresis.

[31]  A. Görg,et al.  Two‐dimensional polyacrylamide gel electrophoresis with immobilized pH gradients in the first dimensin (IPG‐Dalt): The state of the art and the controversy of vertical versus horizontal systems , 1995, Electrophoresis.

[32]  M R Wilkins,et al.  Rapid protein identification using N-terminal "sequence tag" and amino acid analysis. , 1996, Biochemical and biophysical research communications.

[33]  J. Martig,et al.  Evidence for autosomal recessive inheritance of a major gene for bovine dilated cardiomyopathy. , 1998, Journal of animal science.

[34]  M R Wilkins,et al.  Large-scale amino-acid analysis for proteome studies. , 1996, Journal of chromatography. A.

[35]  M. Salpeter,et al.  Agonist-induced myopathy at the neuromuscular junction is mediated by calcium , 1979, The Journal of cell biology.