Creatine kinase binds more firmly to the M-band of rabbit skeletal muscle myofibrils in the presence of its substrates
Language English Country Netherlands Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Adenosine Triphosphate metabolism MeSH
- Models, Biological MeSH
- Fluorescent Dyes chemistry MeSH
- Photobleaching MeSH
- Connectin MeSH
- Muscle Fibers, Skeletal metabolism MeSH
- Muscle, Skeletal metabolism MeSH
- Rabbits MeSH
- Creatine Kinase, MM Form metabolism MeSH
- Muscle Proteins metabolism MeSH
- Protein Binding MeSH
- Animals MeSH
- Check Tag
- Rabbits MeSH
- Male MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Adenosine Triphosphate MeSH
- Fluorescent Dyes MeSH
- Connectin MeSH
- Creatine Kinase, MM Form MeSH
- Muscle Proteins MeSH
Creatine kinase (CK) (E.C. 2.7.3.2) buffers cellular ATP concentration during fluctuating ATP turnover. Muscle cytosolic CK isoform interacts with various subcellular structures where it is functionally coupled with relevant ATPases. However, how this interaction affects its activity is not known. We have therefore studied the interaction of CK with myofibrils and the role of different conformational states of CK molecule induced by ATP, phosphocreatine, ADP and the ATP-creatine pair. Purified rabbit psoas myofibrils with CK specific activity of 0.4+/-0.02 IU/mg were used. The exchange rates between the myofibrillar M-band and its surroundings were measured with fluorofore conjugated CK (IAF) by the Fluorescence Lost in Photobleaching (FLIP) method within a very narrow pH range 7.1-7.15. For CK-IAF without docked substrates, the time derivative of the initial loss of the fluorescent signal within the M-band equalled -3.26 at the fifth second and the decrease reached 82% by the 67th second. For CK-IAF with added substrates, the derivatives fell into the range of -0.95 to -1.30, with respective decreases from 16 to 46% at the 67th second. The results show that the substrates slowed down the exchange rate. This indicates that the strength of the bond between CK and the M-band of myofibrils increased.
Department of Physiology Faculty of Science Charles University Prague Prague 2 128 00 Czech Republic
See more in PubMed
Physiol Res. 2003;52(5):579-85 PubMed
Exp Physiol. 1999 Jul;84(4):651-63 PubMed
Biophys J. 1988 Jun;53(6):935-46 PubMed
Biochem J. 2002 May 1;363(Pt 3):785-92 PubMed
Biochim Biophys Acta. 1971 Aug 6;245(1):259-62 PubMed
Am J Physiol. 1987 Sep;253(3 Pt 1):C444-55 PubMed
J Appl Physiol (1985). 1990 Sep;69(3):968-72 PubMed
Biochim Biophys Acta. 2006 Mar;1760(3):364-71 PubMed
J Cell Sci. 2002 Dec 15;115(Pt 24):4925-36 PubMed
J Muscle Res Cell Motil. 2005;26(6-8):375-9 PubMed
Biochemistry. 2002 Nov 26;41(47):13861-7 PubMed
Mol Cell Biochem. 1994 Apr-May;133-134:125-44 PubMed
Exp Physiol. 2003 Jan;88(1):1-6 PubMed
Biochem J. 1992 Jan 1;281 ( Pt 1):21-40 PubMed
J Cell Biol. 2000 Jun 12;149(6):1225-34 PubMed
J Biol Chem. 1984 Apr 25;259(8):5238-46 PubMed
Physiol Res. 2002;51(1):35-41 PubMed
J Mol Biol. 2003 Sep 26;332(4):877-87 PubMed
J Cell Sci. 1998 May;111 ( Pt 9):1207-16 PubMed
Biochim Biophys Acta. 1996 Jun 13;1274(3):81-8 PubMed
Biophys J. 1995 Oct;69(4):1246-58 PubMed
Biochemistry. 1999 Jun 29;38(26):8492-500 PubMed
Desmin Knock-Out Cardiomyopathy: A Heart on the Verge of Metabolic Crisis
