Comparison of two‐dimensional electrophoresis patterns of heat shock protein Hsp27 species in normal and cardiomyopathic hearts

Heat shock protein Hsp27 occurs in a complex pattern in human myocardial tissue. Normal and failing explanted human heart from patients with dilated cardiomyopathy (DCM) or ischemic heart failure (IHF), respectively, were analyzed by high resolution two‐dimensional electrophoresis (23×30 cm) and Hsp27 immunostaining. Twelve Hsp27 spots in DCM samples were significantly altered in intensity and ten of these were significantly changed in IHF. Four spots (h1, h2, h4, h5) in DCM samples and three spots (h2, h4, h5) in IHF at a molecular mass of 28 kDa were decreased in intensity. In this study, investigating left ventricles of human myocardium, spot h4 was only detected in normal heart samples. On the other hand, spots with a lower molecular mass of 27 kDa (h14, h15, h17, h20, h21) and 22—23 kDa (46, h47, h50) increased in intensity in failing hearts, suggesting that some form of Hsp27 degradation occurs during heart failure.

[1]  W. Schaper,et al.  Expression of heat shock proteins in the normal and stunned porcine myocardium. , 1993, Cardiovascular research.

[2]  J. Landry,et al.  Modulation of actin dynamics during stress and physiological stimulation by a signaling pathway involving p38 MAP kinase and heat-shock protein 27. , 1995, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[3]  D. Mann,et al.  Differential expression of heat shock proteins in normal and failing human hearts. , 1998, Journal of molecular and cellular cardiology.

[4]  David Stokoe,et al.  Identification of MAPKAP kinase 2 as a major enzyme responsible for the phosphorylation of the small mammalian heat shock proteins , 1992, FEBS letters.

[5]  H. Bielka,et al.  Cell-free phosphorylation of the murine small heat-shock protein hsp25 by an endogenous kinase from Ehrlich ascites tumor cells. , 1992, Biochimica et biophysica acta.

[6]  B. Geiger,et al.  A 25-kD inhibitor of actin polymerization is a low molecular mass heat shock protein , 1991, The Journal of cell biology.

[7]  W. Welch,et al.  Characterization and purification of the small 28,000-dalton mammalian heat shock protein. , 1987, The Journal of biological chemistry.

[8]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[9]  W. Welch Phorbol ester, calcium ionophore, or serum added to quiescent rat embryo fibroblast cells all result in the elevated phosphorylation of two 28,000-dalton mammalian stress proteins. , 1985, The Journal of biological chemistry.

[10]  Eckart Fleck,et al.  Protein composition of the human heart: The construction of a myocardial two‐dimensional electrophoresis database , 1994, Electrophoresis.

[11]  E. Müller,et al.  Identification of human myocardial proteins separated by two‐dimensional electrophoresis with matrix‐assisted laser desorption/ionization mass spectrometry , 1996, Electrophoresis.

[12]  J. Behlke,et al.  Phosphorylation and supramolecular organization of murine small heat shock protein HSP25 abolish its actin polymerization-inhibiting activity. , 1994, The Journal of biological chemistry.

[13]  V. Erdmann,et al.  Identification of the phosphorylation sites of the murine small heat shock protein hsp25. , 1991, The Journal of biological chemistry.

[14]  K. Schulze-Osthoff,et al.  Small Stress Proteins as Novel Regulators of Apoptosis , 1996, The Journal of Biological Chemistry.

[15]  M. W. Pandit,et al.  Correlation between stability of a protein and its dipeptide composition: a novel approach for predicting in vivo stability of a protein from its primary sequence. , 1990, Protein engineering.

[16]  P. Allen,et al.  Changes in Myofibrillar Content and Mg‐ATPase Activity in Ventricular Tissues From Patients with Heart Failure Caused by Coronary Artery Disease, Cardiomyopathy, or Mitral Valve Insufficiency , 1988, Circulation research.

[17]  L. Brunton,et al.  Small heat shock proteins and protection against ischemic injury in cardiac myocytes. , 1997, Circulation.

[18]  J. S. Janicki,et al.  Myocardial fibrosis: functional significance and regulatory factors. , 1993, Cardiovascular research.

[19]  E. Müller,et al.  Identification and characterization of heat shock protein 27 protein species in human myocardial two‐dimensional electrophoresis patterns , 1997, Electrophoresis.

[20]  G. Stein,et al.  Sequence and organization of genes encoding the human 27 kDa heat shock protein. , 1986, Nucleic acids research.

[21]  W. Welch,et al.  Interleukin 1 and tumour necrosis factor increase phosphorylation of the small heat shock protein , 1989, FEBS letters.

[22]  R. Klemenz,et al.  Abundance and location of the small heat shock proteins HSP25 and alphaB-crystallin in rat and human heart. , 1997, Circulation.

[23]  M. Gaestel,et al.  Development and tissue-specific distribution of mouse small heat shock protein hsp25. , 1993, Developmental genetics.

[24]  M. Gaestel,et al.  Dephosphorylation of the small heat shock protein hsp25 by calcium/calmodulin-dependent (type 2B) protein phosphatase. , 1992, The Journal of biological chemistry.

[25]  K. Kato,et al.  Copurification of small heat shock protein with alpha B crystallin from human skeletal muscle. , 1992, The Journal of biological chemistry.

[26]  Joachim Klose,et al.  Two‐dimensional electrophoresis of proteins: An updated protocol and implications for a functional analysis of the genome , 1995, Electrophoresis.