Phosphotransfer dynamics in skeletal muscle from creatine kinase gene-deleted mice

Research output: Contribution to journalArticle

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Abstract

To assess the significance of energy supply routes in cellular energetic homeostasis, net phosphoryl fluxes catalyzed by creatine kinase (CK), adenylate kinase (AK) and glycolytic enzymes were quantified using 18O-phosphoryl labeling. Diaphragm muscle from double M-CK/ScCKmit knockout mice exhibited virtually no CK-catalyzed phosphotransfer. Deletion of the cytosolic M-CK reduced CK-catalyzed phosphotransfer by 20%, while the absence of the mitochondrial ScCKmit isoform did not affect creatine phosphate metabolic flux. Contribution of the AK-catalyzed phosphotransfer to total cellular ATP turnover was 15.0, 17.2, 20.2 and 28.0% in wild type, ScCKmit, M-CK and M-CK/ScCKmit deficient muscles, respectively. Glycolytic phosphotransfer, assessed by G-6-P 18O-phosphoryl labeling, was elevated by 32 and 65% in M-CK and M-CK/ScCKmit deficient muscles, respectively. Inhibition of glyceraldehyde 3-phosphate dehydrogenase (GAPDH)/phosphoglycerate kinase (PGK) in CK deficient muscles abolished inorganic phosphate compartmentation and redirected high-energy phosphoryl flux through the AK network. Under such conditions, AK phosphotransfer rate was equal to 86% of the total cellular ATP turnover concomitant with almost normal muscle performance. This indicates that near-equilibrium glycolytic phosphotransfer reactions catalyzed by the GAPDH/PGK support a significant portion of the high-energy phosphoryl transfer in CK deficient muscles. However, CK deficient muscles displayed aberrant ATPase-ATPsynthase communication along with lower energetic efficiency (P/O ratio), and were more sensitive to metabolic stress induced by chemical hypoxia. Thus, redistribution of phosphotransfer through glycolytic and AK networks contributes to energetic homeostasis in muscles under genetic and metabolic stress complementing loss of CK function.

Original languageEnglish (US)
Pages (from-to)13-27
Number of pages15
JournalMolecular and Cellular Biochemistry
Volume256-257
Issue number1-2
StatePublished - Jan 2004

Fingerprint

MM Form Creatine Kinase
Creatine Kinase
Muscle
Skeletal Muscle
Adenylate Kinase
Genes
Muscles
Phosphoglycerate Kinase
Glyceraldehyde-3-Phosphate Dehydrogenases
Physiological Stress
Fluxes
Labeling
Adenosine Triphosphate
Homeostasis
Phosphocreatine
Diaphragms
Energy Transfer
Energy transfer
Diaphragm
Adenosine Triphosphatases

Keywords

  • O-phosphoryl exchange
  • Adenylate kinase
  • Energy metabolism
  • Gene knockout
  • Glycolysis
  • Metabolic networks

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Phosphotransfer dynamics in skeletal muscle from creatine kinase gene-deleted mice. / Dzeja, Petras P; Terzic, Andre; Wieringa, Bé.

In: Molecular and Cellular Biochemistry, Vol. 256-257, No. 1-2, 01.2004, p. 13-27.

Research output: Contribution to journalArticle

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