Basic Science and Experimental Studies
Proteomic Analysis Reveals Significant Alternations of Cardiac Small Heat Shock Protein Expression in Congestive Heart Failure

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Abstract

Background

Because congestive heart failure (CHF) is a complex syndrome with many different underlying mechanisms of worsening of heart function, it is important to recognize the global alternations in protein expression associated with the processes of CHF.

Methods and Results

The purpose of our study was to use a proteomic approach to investigate global alternations in protein expression in tachycardia induced CHF dogs. We compared the 2-dimensional electrophoresis protein patterns of left ventricular samples from the normal with those from failing myocardium. Differentially expressed cardiac proteins showed approximately 500 cardiac protein spots. A total of 20 spots (14 increased, 6 decreased) was altered in CHF, whereas the more distinguishably increased spots in CHF were identified by using mass spectrometry as alpha B crystallin, heat shock protein (HSP) 27, and HSP20, which maintain both the morphologic and functional integrity of the cardiomyocytes and increase tolerance against various types of stress. Because phosphorylation is one of the most important posttranslational modifications, we evaluated whether or not the overexpressed small HSPs were phosphorylated in CHF. Phosphoprotein staining and Western blotting demonstrated that the phosphorylation of alpha B crystallin at serine (Ser)-59 site and of HSP27 at both Ser-78 and Ser-82 sites increased in CHF.

Conclusion

Proteomics studies can provide new insights into molecular mechanisms in CHF and phosphorylated small HSPs may be involved in preventing cardiac dysfunction.

Section snippets

Animal Preparation

All animal experiments were conducted according to the Guide for the Care and Use of Laboratory Animals (Department of Health and Human Services, National Institutes of Health, Publication No. 86-23) and to the Guidelines for Animal Experimentation at the Animal Research Committee of Shiga University of Medical Science. CHF was induced by rapid right ventricular pacing (240 ppm, 28 days) in 6 beagle dogs (CHF group), and a thermodilution catheter was implanted for measurement of cardiac output

Cardiohemodynamics and Blood Analysis

The results of cardiohemodynamics and blood analysis are summarized in Table 1. After 4 weeks of pacing, cardiac output significantly decreased and both LV end-diastolic pressure and pulmonary capillary wedge pressure increased compared to the normal group. LV end-diastolic dimensions increased and LV dP/dT max and LV% fractional shortening decreased compared with the normal group. Plasma norepinephrine levels were significantly higher in the CHF group than in the normal group. These data

Discussion

Using proteomic analysis, we first carried out screening of the protein changes in our paced dog model. Quantitative computer analysis of differentially expressed cardiac proteins showed as many as 500 cardiac protein spots. A total of 20 spots (14 increased, 6 decreased) was altered in CHF and 9 more distinguishably changed protein spots (6 increased, 3 decreased) were identified by using MALDI-TOF MS. The highly expressed spots in CHF were alpha B crystallin, HSP27, and HSP20, which maintain

Acknowledgments

We would like to thank Akiko Yoshii (Department of Cardiovascular and Respiratory Medicine, Shiga University of Medical Science, Shiga, Japan), Noboru Urushiyama, and Masafumi Suzaki (Central Research Laboratory, Shiga University of Medical Science, Shiga, Japan) for their technical assistance.

References (38)

  • H.E. Hoover et al.

    Alpha B crystallin gene induction and phosphorylation by MKK6-activated p38

    J Biol Chem

    (2000)
  • J.J. Hwang et al.

    Genomics and the pathophysiology of heart failure

    Curr Cardiol Rep

    (2001)
  • M. Fujii et al.

    Bradykinin improves left ventricular diastolic function under long-term angiotensin-converting enzyme inhibition in heart failure

    Hypertension

    (2002)
  • E. McGregor et al.

    Proteomics of heart disease

    Hum Mol Genet

    (2003)
  • D.K. Arrell et al.

    Cardiovascular proteomics: evolution and potential

    Circ Res

    (2001)
  • M.B. Yaffe et al.

    Signal transduction. Grabbing phosphoproteins

    Nature

    (1999)
  • L.E. Morrison et al.

    Mimicking phosphorylation of alpha B crystallin on serine-59 is necessary and sufficient to provide maximal protection of cardiac myocytes from apoptosis

    Circ Res

    (2003)
  • A. Wada et al.

    Cardiorenal and neurohumoral effects of endogenous atrial natriuretic peptide in dogs with severe congestive heart failure using a specific antagonist for guanylate cyclase-coupled receptors

    Circulation

    (1994)
  • A. Wada et al.

    Effects of a specific endothelin-converting enzyme inhibitor on cardiac, renal, and neurohumoral functions in congestive heart failure: comparison of effects with those of endothelin A receptor antagonism

    Circulation

    (1999)
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    Supported by a grant-in-aid for Scientific Research (C) and a grant-in-aid for Encouragement of Young Scientists from the Minister of Education, Culture, Sports, Science and Technology of Japan.

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