Skip to main content
SpringerLink
Log in

Coupling of creatine kinase to glycolytic enzymes at the sarcomeric I-band of skeletal muscle: a biochemical study in situ

  • Published:
Journal of Muscle Research & Cell Motility Aims and scope Submit manuscript

Abstract

The specific interaction of muscle type creatine-kinase (MM-CK) with the myofibrillar M-line was demonstrated by exchanging endogenous MM-CK with an excess of fluorescently labeled MM-CK in situ, using chemically skinned skeletal muscle fibers and confocal microscopy. No binding of labeled MM-CK was noticed at the I-band of skinned fibers, where the enzyme is additionally located in vivo, as shown earlier by immunofluorescence staining of cryosections of intact muscle. However, when rhodamine-labeled MM-CK was diffused into skinned fibers that had been preincubated with phosphofructokinase (PFK), a glycolytic enzyme known to bind to actin, a striking in vivo-like interaction of Rh-MM-CK with the I-band was found, presumably mediated by binding of Rh-MM-CK to the glycolytic enzyme. Aldolase, another actin-binding glycolytic enzyme was also able to bind Rh-MM-CK to the I-band, but formation of the complex occurred preferably at long sarcomere length (>3.0 μm). Neither pyruvate kinase, although known for its binding to actin, nor phosphoglycerate kinase (PGK), not directly interacting with the I-band itself, did mediate I-band targeting of MM-CK. Anchoring of MM-CK to the I-band via PFK, but not so via aldolase, was strongly pH-dependent and occurred below pH 7.0. Labeling performed at different sarcomere length indicated that the PFK/MM-CK complex bound to thin filaments of the I-band, but not within the actomyosin overlap zones. The physiological consequences of the structural interaction of MM-CK with PFK at the I-band is discussed with respect to functional coupling of MM-CK to glycolysis, metabolic regulation and channeling in multi-enzyme complexes. The in situ binding assay with skinned skeletal muscle fibers described here represents a useful method for further studies of specific protein–protein interactions in a structurally intact contractile system under various precisely controlled conditions.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Adams V, Griffin L, Towbin J, Gelb B, Worley K and McCabe ER (1991) Porin interaction with hexokinase and glycerol kinase: metabolic microcompartmentation at the outer mitochondrial membrane. Biochem Med Metab Biol 45: 271–291.

    Google Scholar 

  • Arnold H and Pette D (1968) Binding of glycolytic enzymes to structure proteins of the muscle. Eur J Biochem 6(2): 163–171.

    Google Scholar 

  • Arnold H, Henning R and Pette D (1971) Quantitative comparison of the binding of various glycolytic enzymes to F-actin and the interaction of aldolase with G-actin. Eur J Biochem 22: 121–126.

    Google Scholar 

  • Arrio-Dupont M, Bechet JJ and D'Albis A (1992) A model system of of coupled activity of co-immobilized creatine kinase and myosin. Eur J Biochem 207: 951–955.

    Google Scholar 

  • Brenner B (1983) Technique for stabilizing the striation pattern in maximally calcium-activated skinned rabbit psoas fibers. Biophys J 41: 99–102.

    Google Scholar 

  • Bronstein WW and Knull HR (1981) Interaction of muscle glycolytic enzymes with thin filament proteins. Can J Biochem 59: 494–499.

    Google Scholar 

  • Brumback RA, Gerst JW and Knull HR (1983) High energy phosphate depletion in a model of defective muscle glycolysis. Mus Ner 6: 52–55.

    Google Scholar 

  • Clarke FM and Masters CJ (1975) On the association of glycolytic enzymes with structural proteins of skeletal muscle. Biochim Biophys Acta 381: 37–46.

    Google Scholar 

  • Dillon PF and Clark JF (1990) The theory of diazymes and functional coupling of pyruvate kinase and creatine kinase. J Theor Biol 143: 275–284.

    Google Scholar 

  • DÄlken G, Leisner E and Pette D (1975) Immunofluorescent localization of glycogenolytic and glycolytic enzyme proteins and of malate dehydrogenase isozymes in cross-striated skeletal muscle and heart of the rabbit. Histochemistry 43: 113–121.

    Google Scholar 

  • Eppenberger HM, Perriard JC, Rosenberg UB and Strehler EE (1981) The Mr 165,000 M-protein myomesin. A specific protein of cross-striated muscle cells. J Cell Biol 89: 185–193.

    Google Scholar 

  • Fabiato A and Fabiato F (1975) Dependence of the contractile activation of skinned cardiac muscle cells on the sarcomere length. Nature 256: 54–56.

    Google Scholar 

  • Foucault G, Vacher M, Merkulova T, Keller A and Arrio-Dupont M (1999) Presence of enolase in the M-band of skeletal muscle and possible indirect interaction with the cytosolic isoform of creatine kinase. Biochem J 338: 115–121.

    Google Scholar 

  • Freydina NA, Shpagina MD and Podlubnaya ZA (1986) Localization of binding sites of F-protein (phosphofructokinase) on the myosin molecule. J Muscle Res Cell Motil 7: 481–490.

    Google Scholar 

  • Gerlach G and Hofer HW (1986) Interaction of immobilized phosphofructokinase with soluble muscle proteins. Biochim Biophys Acta 881: 398–404.

    Google Scholar 

  • Goding JW (1976) Conjugation of antibodies with fluorochromes: modification to the standard methods. J Immunol Methods 13: 215–226.

    Google Scholar 

  • Gordon AM, Huxley AF and Julian FJ (1966) The variation in isometric tension with sarcomere length in vertebrate muscle fibers. J Physiol (Lond) 184: 170–192.

    Google Scholar 

  • Grove BK, Kurer V, Lehner Ch, Doetschman TC, Perriard JC and Eppenberger HM (1984) A new 185,000-dalton skeletal muscle protein detected by monoclonal antibodies. J Cell Biol 98: 518–524.

    Google Scholar 

  • Hornemann T, Stolz M and Wallimann T (2000) Isoenzyme-specific interaction of muscle-type creatine kinase with the sarcomeric M-line is mediated by NH(2)-terminal lysine charge-clamps. J Cell Biol 149(6): 1225–1234.

    Google Scholar 

  • Han JW, Thieleczek R, Varsanyi M and Heilmeyer LMG (1992) Compartmentalized ATP synthesis in skeletal muscle triads. Biochemistry 31: 377–384.

    Google Scholar 

  • Kraft T, Chalovich JM, Yu LC and Brenner B (1995a) Parallel inhibition of active force and relaxed fiber stiffness by caldesmon fragments at physiological ionic strength and temperature conditions: additional evidence that weak cross-bridge binding to actin is an essential intermediate for force generation. Biophys J 68: 2404–2418.

    Google Scholar 

  • Kraft T, Messerli M, Rothen-Rutishauser B, Perriard J-C, Wallimann T and Brenner B (1995b) Equilibration and exchange of fluorescently labeled molecules in skinned skeletal muscle fibers visualized by confocal microscopy. Biophys J 69: 1246–1258.

    Google Scholar 

  • Kraft T, Messerli M, Rutishauser B, Wallimann T, Perriard J-C, Chalovich JM and Brenner B (1995c) Equilibration and exchange of fluorescently labeled molecules in skeletal muscle fibers studied using confocal microscopy. Biophys J 68: 371s.

    Google Scholar 

  • Kraft T, Nier V, Brenner B and Wallimann T (1996) Binding of creatine kinase to the I-band of skinned skeletal muscle fibers is mediated by glycolytic enzymes: an in situ biochemical approach. Biophys J 70: A292.

    Google Scholar 

  • Kuprianov VV, Seppet EK, Emelin IV and Saks VA (1980) Phosphocreatine production coupled to the glycolytic reactions in the cytosol of cardiac cells. Biochim Biophys Acta 592: 197–210.

    Google Scholar 

  • Lamsen SE, Belknap JK, Sampson KE and Knull HR (1990) Hexokinase redistribution in vivo. Biochim Biophys Acta 1034: 118–121.

    Google Scholar 

  • Liou RS and Anderson S (1980) Activation of rabbit muscle phosphofructokinase by F-actin and reconstituted thin filaments. Biochemistry 19: 2684–2688.

    Google Scholar 

  • Masters CJ (1981) Interactions between soluble enzymes and subcellular structure. Crit Rev Biochem II: 105–144.

    Google Scholar 

  • Maughan D and Wegner E (1989) On the organization and diffusion of glycolytic enzymes in skeletal muscle. In: Paul R, Elzinga G and Yamada K (eds) Progress in Clinical and Biological Research. Vol. 315, Muscle Energetics, pp. 137–147.

  • Májean C, Pons F, Benyamin Y and Roustan C (1989) Antigenic probes locate binding site for the glycolytic enzymes GPDH, aldolase and phosphofructokinase on the actin monomer in microfilaments. Biochem J 264: 671–677.

    Google Scholar 

  • Ovadi J (1995) Cell Architecture and Metabolic Channeling. Molecular Biology Intelligence Unit, Springer, R.G. Landes Co. Austin, Texas.

    Google Scholar 

  • Ovadi J and Orosz F (1997) A new concept for control of glycolysis. In: Agius L and Sherratt HSA (eds) Channelling in Intermediary Metabolism. (pp. 237–268) Portland Press, London.

    Google Scholar 

  • Parmeggiani A, Luft JH, Love DS and Krebs EG (1966) Crystallization and properties of rabbit skeletal muscle phosphofructokinase. J Biol Chem 241: 4625–4637.

    Google Scholar 

  • Parra J and Pette D (1995) Effect of low-frequency stimulation on soluble and structure-bound activities of hexokinase and phosphofructokinase in rat fast-twitch muscle. Biochim Biophys Acta 1251: 154–160.

    Google Scholar 

  • Paul RJ, Hardin CD, Raeymaekers L, Wuytack F and Casteels R (1989) An endogenous glycolytic cascade can preferentially support Ca2+-uptake in smooth muscle plasmamembrane vesicles. FASEB J 3: 2290–2301.

    Google Scholar 

  • Pette D (1975) Some aspects of supramolecular organization of glycogenolytic and glycolytic enzymes in muscle. Acta histochem Suppl. XIV: 47–68.

    Google Scholar 

  • Podlubnaya ZA (1992) Enzymes in thick filaments of vertebrate striated muscles. Biochemistry-Russia 57: 1239–1259.

    Google Scholar 

  • Roberts SJ and Somero GN (1987) Binding of phosphofructokinase to filamentous actin. Biochemistry 26: 3437–3442.

    Google Scholar 

  • Roberts SJ, Lowery MS and Somero GN (1988) Regulation of binding of phosphofructokinase to myofibrils in the red and white muscle of the barred sand bass, Paralabrax nebulifer (Serranidae). J Exp Biol 137: 13–27.

    Google Scholar 

  • Rossi AM, Eppenberger HM, Volpe P, Cotrufo R and Wallimann T (1990) Muscle-type MM-creatine kinase is specifically bound to sarcoplasmic reticulum and can support Ca2+-uptake and regulate local ATP/ADP ratios. J Biol Chem 265: 5258–5266.

    Google Scholar 

  • Rüdel R and Taylor SR (1971) Striated muscle fibers: facilitation of contraction at short muscle lengths by caffeine. Science 172: 387–388.

    Google Scholar 

  • Sata M, Sugiura S, Yamashita H, Momomura S and Serizawa T (1996) Coupling between myosin ATPase cycle and creatine kinase facilitates cardiac actomyosin sliding in vitro. Circulation 93: 310–317.

    Google Scholar 

  • Schoenberg M and Podolsky RJ (1972) Length-force relation of calcium activated muscle fibers. Science 176: 52–54.

    Google Scholar 

  • Scopes RK (1973) Studies with a reconstituted muscle glycolytic system. Biochem J 134: 197–208.

    Google Scholar 

  • Sears PR and Dillon PF (1999) Differential coupling of smooth and skeletal muscle pyruvate kinase. Biochemistry 9: 14881–14886.

    Google Scholar 

  • Sigel P and Pette D (1969) Intracellular localization of glycogenolytic and glycolytic enzymes in white and red rabbit skeletal muscle. J Histochem Cytochem 17: 225–237.

    Google Scholar 

  • Somero GN and Hand SC (1990) Protein assembly and metabolic regulation: physiological and evolutionary perspectives. Physiol Zool 63: 443–471.

    Google Scholar 

  • Srere PA and Knull HR (1998) Location — location — location. TIBS 23: 319–320.

    Google Scholar 

  • Stolz M, Kraft T and Wallimann T (1998) The isoenzyme-diagnostic regions of muscle-type creatine kinase, the M-260 and M-300 box, are not responsible for its myofibrillar M-line binding. Europ J Cell Biol 77: 1–9.

    Google Scholar 

  • Stolz M and Wallimann T (1998) Myofibrillar interaction of cytosolic creatine kinase (CK) isoenzymes: allocation of N-terminal binding epitope in MM-CK and BB-CK. J Cell Sci 111: 1207–1216.

    Google Scholar 

  • The TH and Feltkamp TEW (1970a) Conjugation of fluorescein isothiocyanate to antibodies. I. Experiments on the conditions of conjugation. Immunology 18: 865–873.

    Google Scholar 

  • The TH and Feltkamp TEW (1970b) Conjugation of fluorescein isothiocyanate to antibodies. II. A reproducible method. Immunology 18: 875–881.

    Google Scholar 

  • Turner D, Wallimann T and Eppenberger HM (1973) A protein that binds specifically to the M-line of skeletal muscle is identified as the muscle form of creatine kinase. Proc Natl Acad Sci USA 70: 702–706.

    Google Scholar 

  • Uyeda K and Racker E (1965) Regulatory mechanisms in carbohydrate metabolism. VII. Hexokinase and phosphofructokinase. J Biol Chem 240: 4682–4688.

    Google Scholar 

  • Van Waarde A, Van den Thillart G, Erkelens C, Addink A and Lugtenburg J (1990) Functional coupling of glycolysis and phosphocreatine utilization in anoxic fish muscle. An in vivo 31P NMR study. J Biol Chem 265: 914–923.

    Google Scholar 

  • Ventura-Clapier R, Mekhfi H and Vassort G (1987) Role of creatine kinase in force development in chemically skinned rat cardiac muscle. J Gen Physiol 89: 815–837.

    Google Scholar 

  • Ventura-Clapier R, Veksler V and Hoerter JA (1994) Myofibrillar creatine kinase and cardiac contraction. Mol Cell Biochem 133/134: 125–144.

    Google Scholar 

  • Wallimann T, Turner DC and Eppenberger HM (1975) Creatine kinase and M-line structure. FEBS Proc 31: 119–124.

    Google Scholar 

  • Wallimann T, Moser H and Eppenberger H (1983) Isoenzyme-specific localization of M-line bound creatine kinase in myogenic cells. J Muscle Res Cell Mot 4: 429–441.

    Google Scholar 

  • Wallimann T, Schlösser T and Eppenberger HM (1984) Function of M-line-bound CK as intramyofibrillar ATP regenerator at the receiving end of the phosphorylcreatine shuttle in muscle. J Biol Chem 259: 5238–5246.

    Google Scholar 

  • Wallimann T and Eppenberger HM (1985) Localization and function of M-line-bound creatine kinase. M-band model and creatine phosphate shuttle. Cell Muscle Motil 6: 239–285. Review.

    Google Scholar 

  • Wallimann T, Wyss M, Brdiczka D, Nicolay K and Eppenberger HM (1992) Intracellular compartmentation, structure and function of creatine kinase isoenzymes in tissues with high and fluctuating energy demands: the ‘phosphocreatine circuit’ for cellular energy homeostasis. Biochem J 281: 21–40.

    Google Scholar 

  • Wallimann T and Hemmer W (1994) Creatine kinase in non-muscle tissues and cells. Mol Cell Biochem 133–134: 193–220.

    Google Scholar 

  • Walsh TP, Winzor DJ, Clarke FM, Masters CJ and Morton DJ (1980) Binding of aldolase to actin-containing filaments. Evidence of interaction with the regulatory proteins of skeletal muscle. Biochem J 186: 89–98.

    Google Scholar 

  • Walsh TP, Masters SJ, Morton DJ and Clarke FM (1981) The reversible binding of glycolytic enzymes in ovine skeletal muscle in response to tetanic stimulation. Biochim Biophys Acta 675: 29–39.

    Google Scholar 

  • Wegener G (1990) Elite invertebrate athletes: flight in insects, its metabolic requirements and regulation and its effects on life span. In: Nazar K, Terjung RL, Kaciuba-Uscilko H and Budohoski L (eds) International Perspectives in Exercise Physiology, Human Kinetics Publ., Champaign, Illinois, pp 83–87.

    Google Scholar 

  • Wegmann E, Zanolla E, Eppenberger HM and Wallimann T (1992) In situ compartmentation of creatine kinase in intact sarcomeric muscle: the acto-myosin overlap zone as a molecular sieve. J Muscle Res Cell Mot 13: 420–435.

    Google Scholar 

  • Wilson JE (1978) Ambiquitous enzymes: variation in intracellular distribution a regulatory mechanism. TIBS 3: 124–125.

    Google Scholar 

  • Wojtas K, Slepecky N, von Kalm L and Sullivan D (1997) Flight muscle function in Drosophila requires colocalization of glycolytic enzymes. Mol Biol Cell 8: 1665–1675.

    Google Scholar 

  • Yu LC and Brenner B (1989) Structures of actomyosin cross-bridges in relaxed and rigor muscle fibers. Biophys J 55: 441–453.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Molekular- und Zellphysiologie, Medizinische Hochschule Hannover, D-30625, Hannover, Germany

    Theresia Kraft & V. Nier

  2. Swiss Federal Institute of Technology, Institute of Cell Biology, ETH Zürich, Hönggerberg, CH-8093, Zürich, Switzerland

    T. Hornemann, M. Stolz & T. Wallimann

Authors
  1. Theresia Kraft
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T. Hornemann
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. Stolz
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. V. Nier
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. T. Wallimann
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kraft, T., Hornemann, T., Stolz, M. et al. Coupling of creatine kinase to glycolytic enzymes at the sarcomeric I-band of skeletal muscle: a biochemical study in situ. J Muscle Res Cell Motil 21, 691–703 (2000). https://doi.org/10.1023/A:1005623002979

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1005623002979

Keywords

Search

Navigation