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Non-targeted metabolomics analysis of cardiac Muscle Ring Finger-1 (MuRF1), MuRF2, and MuRF3 in vivo reveals novel and redundant metabolic changes

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Abstract

The muscle-specific ubiquitin ligases MuRF1, MuRF2, MuRF3 have been reported to have overlapping substrate specificities, interacting with each other as well as proteins involved in metabolism and cardiac function. In the heart, all three MuRF family proteins have proven critical to cardiac responses to ischemia and heart failure. The non-targeted metabolomics analysis of MuRF1−/−, MuRF2−/−, and MuRF3−/− hearts was initiated to investigate the hypothesis that MuRF1, MuRF2, and MuRF3 have a similarly altered metabolome, representing alterations in overlapping metabolic processes. Ventricular tissue was flash frozen and quantitatively analyzed by GC/MS using a library built upon the Fiehn GC/MS Metabolomics RTL Library. Non-targeted metabolomic analysis identified significant differences (via VIP statistical analysis) in taurine, myoinositol, and stearic acid for the three MuRF−/− phenotypes relative to their matched controls. Moreover, pathway enrichment analysis demonstrated that MuRF1−/− had significant changes in metabolite(s) involved in taurine metabolism and primary acid biosynthesis while MuRF2−/− had changes associated with ascorbic acid/aldarate metabolism (via VIP and t test analysis vs. sibling-matched wildtype controls). By identifying the functional metabolic consequences of MuRF1, MuRF2, and MuRF3 in the intact heart, non-targeted metabolomics analysis discovered common pathways functionally affected by cardiac MuRF family proteins in vivo. These novel metabolomics findings will aid in guiding the molecular studies delineating the mechanisms that MuRF family proteins regulate metabolic pathways. Understanding these mechanism is an important key to understanding MuRF family proteins’ protective effects on the heart during cardiac disease.

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Abbreviations

E3:

Ubiquitin ligase

MuRF1(2,3):

Muscle ring finger-1(2,3)

VIP:

Variable importance in projection

PCA:

Principal components analysis

PLS-DA:

Partial least squares discriminant analysis

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Acknowledgments

The authors would like to thank Tim Koves for his guidance and valuable discussion and suggestions for harvesting and preparing heart samples for metabolomics analysis. This work was supported by the National Institutes of Health (R01HL104129 to M.W.), a Jefferson-Pilot Corporation Fellowship (to M.W.), and the Fondation Leducq (to M.W.).

Conflict of interest

The authors report no conflicts of interest.

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Authors and Affiliations

  1. University of North Carolina School of Medicine, Chapel Hill, NC, USA

    Ranjan Banerjee, Megan T. Quintana & Zhongjing Wang

  2. General Hospital of Ningxia Medical University, Yinchuan, Ningxia, People’s Republic of China

    Jun He, Megan T. Quintana & Zhongjing Wang

  3. Department of Pathology & Laboratory Medicine, McAllister Heart Institute, University of North Carolina, 111 Mason Farm Road, MBRB 2340B, Chapel Hill, NC, 27599, USA

    Jun He, Carolyn Spaniel, Megan T. Quintana, Zhongjing Wang & Monte S. Willis

  4. Sarah W. Stedman Nutrition and Metabolism Center, Duke Molecular Physiology Institute, Duke University Medical Center, Durham, NC, USA

    Megan T. Quintana, Zhongjing Wang, James R. Bain, Christopher B. Newgard & Michael J. Muehlbauer

  5. Division of Endocrinology, Metabolism, and Nutrition, Department of Medicine, Duke University Medical Center, Durham, NC, USA

    Megan T. Quintana, Zhongjing Wang, James R. Bain & Christopher B. Newgard

  6. McAllister Heart Institute, University of North Carolina, Chapel Hill, NC, USA

    Megan T. Quintana, Zhongjing Wang & Monte S. Willis

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  1. Ranjan Banerjee
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  2. Jun He
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  9. Monte S. Willis
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Banerjee, R., He, J., Spaniel, C. et al. Non-targeted metabolomics analysis of cardiac Muscle Ring Finger-1 (MuRF1), MuRF2, and MuRF3 in vivo reveals novel and redundant metabolic changes. Metabolomics 11, 312–322 (2015). https://doi.org/10.1007/s11306-014-0695-1

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