title: Mitochondrial creatine kinase isoform expression does not correlate with its mode of action.
abstract: In adult mammalian ventricular tissue, mitochondrial creatine kinase (mi-CK), which is bound to the outer surface of the mitochondrial inner membrane, is functionally coupled to oxidative phosphorylation. This is shown, in saponin-permeabilized rat ventricular fibres, by a shift in the apparent K(m) of mitochondrial respiration for ADP from 300 +/- 56 microM to 111 +/- 40 microM (P < 0.001) on the addition of 25 mM creatine, due to a local accumulation of ADP close to the ATP/ADP translocator. We have found that in atrial fibres, the apparent K(m) for ADP is high, but is not decreased by creatine, suggesting an absence of coupling in this tissue, as has previously been observed in smooth muscle. mi-CK is encoded by two different genes, which are expressed in a tissue-specific manner: the sarcomeric isoform is expressed in ventricular and skeletal muscles, while the ubiquitous isoform is expressed in smooth muscle, brain and other tissues. In order to determine whether a specific function can be attributed to the expression of a specific isoform, we investigated mi-CK mRNA expression by Northern blot analysis. Hybridization with synthetic oligonucleotides specific for each mi-CK isoform showed the expression of only the sarcomeric isoform in rat atria. This result was confirmed by PCR using primers specific for each isoform. In addition, electrophoretic analysis of CK isoforms showed no difference in the octamer/dimer ratio of mi-CK in the atria and ventricles. In atria, unlike the soleus or ventricles, the maximum potential rate of mitochondrial phosphocreatine synthesis was lower than the maximal rate of ATP production by the mitochondria. The total CK/adenylate kinase ratio was also lower in atria than in the other tissues, suggesting a greater contribution of adenylate kinase to adenine nucleotide compartmentation in this tissue. The functional differences between mi CK in the two cardiac tissues seem to imply a specific arrangement of the proteins in the intermembrane space rather than the expression of specific isoforms.
authors.affiliation: U-446 INSERM, Faculté de Pharmacie, Université Paris-Sud, Châtenay-Malabry, France.
authors.forename: K
authors.initials: K
authors.lastname: Anflous
authors.forename: V
authors.initials: V
authors.lastname: Veksler
authors.forename: P
authors.initials: P
authors.lastname: Mateo
authors.forename: F
authors.initials: F
authors.lastname: Samson
authors.forename: V
authors.initials: V
authors.lastname: Saks
authors.forename: R
authors.initials: R
authors.lastname: Ventura-Clapier
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000263:Adenylate Kinase / Q000378:metabolism; D000818:Animals; D015152:Blotting, Northern; D003402:Creatine Kinase / Q000096:biosynthesis* / Q000378:metabolism*; D006325:Heart Atria / Q000201:enzymology / Q000378:metabolism; D006352:Heart Ventricles / Q000201:enzymology / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism; D007527:Isoenzymes; D007700:Kinetics; D008297:Male; D008931:Mitochondria, Muscle / Q000737:chemistry* / Q000201:enzymology*; D018482:Muscle, Skeletal / Q000201:enzymology / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D010101:Oxygen Consumption; D010539:Permeability; D016133:Polymerase Chain Reaction; D051381:Rats; D017208:Rats, Wistar; D013329:Structure-Activity Relationship
chemical_list: D007527:Isoenzymes; D000244:Adenosine Diphosphate; D003402:Creatine Kinase; D000263:Adenylate Kinase