Changes in the mitochondrial proteome from mouse hearts deficient in creatine kinase.
Creatine kinase (CK) is an abundant enzyme, important for maintenance of high-energy phosphate homeostasis in many tissues including heart. Double-knockout CK (DbKO-CK) mice missing both the muscle (MM) and sarcomeric mitochondrial (ScMit) isoforms of CK have recently been studied. Despite a large change in skeletal muscle function in DbKO-CK mice, there is little functional change in the heart. To investigate whether there are specific changes in cardiac mitochondrial proteins associated with the loss of MM- and ScMit-CK isoforms, we have used difference gel electrophoresis (DIGE) to compare mitochondrial proteins from wild-type and DbKO-CK mice. Mass spectrometry fingerprinting was used to identify 40 spots as known mitochondrial proteins. We have discovered that the loss of MM- and ScMit-CK isoforms did not cause large scale changes in heart mitochondrial proteins. The loss of ScMit-CK was readily detected in the DbKO-CK samples. We have also detected a large decrease in the precursor form of aconitase. Furthermore, two mitochondrial protein differences have been found in the parent mouse strains of the DbKO-CK mice.
Kernec
F
F
Laboratory of Functional and Molecular Imaging, National Institute of Neurological Disorders and Stroke, Bethesda, Maryland 20892, USA.
Unlü
M
M
Labeikovsky
W
W
Minden
J S
JS
Koretsky
A P
AP
D000154:Aconitate Hydratase / Q000378:metabolism; D000818:Animals; D002457:Cell Extracts; D003402:Creatine Kinase / Q000235:genetics* / Q000502:physiology*; D052283:Creatine Kinase, MM Form; D052281:Creatine Kinase, Mitochondrial Form; D015180:Electrophoresis, Gel, Two-Dimensional; D007527:Isoenzymes / Q000235:genetics / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008929:Mitochondria, Heart / Q000201:enzymology* / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D002457:Cell Extracts; D007527:Isoenzymes; D020543:Proteome; D003402:Creatine Kinase; D052283:Creatine Kinase, MM Form; D052281:Creatine Kinase, Mitochondrial Form; D000154:Aconitate Hydratase