Summary
Two isoforms of creatine kinase (CK, E.C. 2.7.3.2), the brain type BB-CK and the mitochondrial-bound MiMi-CK, as well as adenylate kinase (myokinase, E.C. 2.7.4.3) were identified in washed spermatozoa from chicken and man by cellulose polyacetate electrophoresis and immunoblots. BB-CK was localized by indirect immunofluorescence staining within the sperm tail but not in the head portion. MiMi-CK is confined to the midpiece region rich in mitochondria and has been localized directly by immunogold staining within the mitochondria. In contrast to chicken, seminal plasma from man was also found to contain considerable amounts of BB-CK. Total creatine content of spermatozoa (8–15mm) and seminal plasma (3.8±0.4m) as well as preliminary experiments with metabolic blockers indicate a dependence of sperm motility on CK and phosphoryl creatine (CP). The presence of two CK isoforms located in different ‘compartments’ of spermatozoa suggests a CP-shuttle in sperm similar to that described for cross-striated muscle.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Arcidiacono, A., Walt, H., Campana, A. &Balerna, M. (1983) The use of percoll gradients for preparation of subpopulations of human spermatozoa.Int. J. Andr. 6, 433–45.
Asseo, PL., Panidis, D., Papadimas, J. &Ikkos, D. (1981) Creatine kinase in human seminal plasma of infertile men: activity and isoenzymes.Israel J. med. Sci. 17, 764–5.
Bessman, S. P. &Geiger, P. J. (1981) Transport of energy in muscle. The phosphorylcreatine shuttle.Science, N.Y. 211, 448–52.
Bishop, D. W. (1962)Spermatozoan Motility. American Association for the Advancement of Science, Washington, D.C., U.S.A. Publication No.72.
Blum, H. E., Deus, G., &Gerok, W. (1983) Mitochondrial creatine kinase from human heart muscle: purification and characterization of the crystallized isoenzyme.J. Biochem. 94, 1247–57.
Buckland, R. B. (1970) Some enzyme activities of chicken spermatozoa.Poult. Sci. 49, 1638–41.
Chapman, J. F., Silverman, L. M., Hall, M. &McGraw, T. (1980) Antibodies to creatine kinase BB in infertile females.Clin. Chem. 26, 1049a.
Cohen, A., Buckingham, M. &Gros, F. (1978) A modified assay procedure for revealing the M-form of CK in cultured muscle cells.Expl Cell Res. 115, 204–7.
Edwards, J. J., Anderson, N. G., Tollaksen, S. L., Eschenbach, A. C. &Guevara, J. (1982) Proteins of human urine. II. Identification by 2d-PAGE of a new candidate marker for prostatic cancer.Clin. Chem. 28, 160–3.
Eggleton, P., Elsden, R. S. &Gough, N. (1943) The estimation of creatine and of diacetyl.Biochem. J. 37, 526–9.
Gonzales-Buitrago, J. M., Miralles, J. M., Munoz, M. H., Meza, S., Alonso, M. T. &Garcia-Diez, L. C. (1980) Seminal plasma creatine kinase activity in fertility studies.Archs Andr. 5, 355–60.
Gornall, A. G., Bardawill, C. J. &David, M. M. (1949) Determination of serum proteins by means of the Biuret reaction.J. biol. Chem. 177, 751–6.
Henderson, A. R. &Nealon, D. A. (1981) Enzyme measurements by mass: an interim review of the clinical efficacy of some mass measurements of prostatic acid phosphatase and the isoenzymes of creatine kinase.Clin. Chem. Acta 115, 9–32.
Jacobus, W. E. &Ingwall, J. S. (eds) (1980)Heart Creatine Kinase. The Integration of Isoenzymes for Energy Distribution. Baltimore, London: Williams and Wilkins.
Jacobus, W. E. &Lehninger, A. L. (1973) CK of rat heart mitochondria.J. biol. Chem. 248, 4803–10.
Kavanagh, J. P. &Darby, C. (1983) Creatine kinase and ATPase in human seminal fluid and prostatic fluid.J. Reprod. Fert. 68, 51–6.
Laemmli, U. K. (1970) Cleavage of structural proteins during assembly of the head of bactriophage T4.Nature, Lond. 277, 680–5.
Lindemann, Ch. B. &Gibbons, I. R. (1975) ATP-induced motility and sliding of filaments in mammalian sperm extracted with Triton X-100.J. Cell Biol. 65, 147–62.
Lowry, O. H., Rosebrough, N. G., Farr, A. L. &Randall, R. J. (1951) Protein measurements with the folin-phenol reagent.J. biol. Chem. 193, 265–75.
Menaché, R., Rubinstein, J., Israeli, S. &Halbrecht, I. (1976) Enzymes and isoenzymes in spermatic fluid and their relationship to spermatozoa concentration.Israel J. Med. Sci. 12, 1353.
Meyer, R. A., Sweeney, H. L. &Kushmerick, M. J. (1984) A simple analysis of the ‘phosphocreatine shuttle’.Am. J. Physiol. 246, C365–77.
Perriard, J. C., Caravatti, M., Perriard, E. &Eppenberger, H. M. (1978) Quantitation of CK isoenzyme transition in differentiating chicken embryonic breast muscle and myogenic cell cultures by immunoadsorption.Archs Biochem. Biophys. 191, 90–100.
Saks, V. A., Rosenstraukh, L. V., Smirnov, V. N. &Chazov, E. I. (1978) Role of creatine phosphokinase in cellular function and metabolism.Can. J. Physiol. Pharmac. 56, 691–706.
Soufir, J. C. (1979) Creatine kinase in human seminal plasma: activity, origin, and isoenzymes.Clin. Chem. 25, 1183–4.
Tombes, R. M., Schackmann, R. W. &Shapiro, B. M. (1984) Sea urchin sperm creatine kinase activity is essential for optimal respiration and motility.J. Cell Biol. 99, 258a.
Tombes, R. T. &Shapiro, B. M. (1985) Metabolite channeling: a phosphorylcreatine shuttle to mediate high energy phosphate transport between sperm mitochondrion and tail.Cell 41, 325–34.
Towbin, H., Staehli, T. &Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications.Proc. natn. Acad. Sci. U.S.A. 76, 4350–4.
Turner, D. C., Maier, V. &Eppenberger, H. M. (1974) CK and aldolase isoenzyme transitions in cultures of chick skeletal muscle cells.Devl Biol. 37, 63–89.
Turner, D. C., Wallimann, T. &Eppenberger, H. M. (1973) A protein that binds specifically to the M-line of skeletal muscle is identified as the muscle form of creatine kinase.Proc. natn. Acad. Sci. U.S.A. 70, 702–5.
Wallimann, T. (1975)Creatinkinase-Isoenzyme und Myofibrillenstruktur. Ph.D. thesis, Eidgenössische Technische Hochschule Zürich, Switzerland.
Wallimann, T., Doetschman, Th. C. &Eppenberger, H. M. (1983a) Novel staining pattern of skeletal muscle M-lines upon incubation with antibodies against MM-CK.J. Cell Biol. 96, 1772–9;98, 785.
Wallimann, T. &Eppenberger, H. M. (1984) Localization and function of M-line-bound CK. M-band model and CP-shuttle. InCell and Muscle Motility, (edited byShay, J. W., Vol. 6. pp. 239–85. New York: Plenum Press.
Wallimann, T., Hardwicke, P. M. D. &Szent-Györgyi, A. G. (1982) Regulatory and essential light-chain interactions in scallop myosin.J. molec. Biol. 156, 153–73.
Wallimann, T., Moser, H. &Eppenberger, H. M. (1983b) Isoenzyme-specific localization of M-line bound CK in myogenic cells.J. Musc. Res. Cell Motility 4, 429–41.
Wallimann, T., Moser, H., Zurbriggen, B. &Eppenberger, H. M. (1984) Creatine kinase (CK) isoenzymes in spermatozoa from chicken and man.Experientia 40, 641.
Wallimann, T., Pelloni, G. W., Turner, D. C. &Eppenberger, H. M. (1978) Monovalent antibodies against MM-creatine kinase remove the M-line from myofibrils.Proc. natn. Acad. Sci. U.S.A. 75, 4296–300.
Wallimann, T., Schlösser, T. &Eppenberger, H. M. (1984) Function of M-line-bound CK as intramyofibrillar ATP regenerator at the receiving end of the phosphoryl-creatine shuttle in muscle.J. biol. Chem. 259, 5238–46.
Wallimann, T., Turner, D. C. &Eppenberger, H. M. (1977) Localization of creatine kinase isoenzymes in myofibrils. I. Chicken skeletal muscle.J. Cell Biol. 75, 297–317.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Wallimann, T., Moser, H., Zurbriggen, B. et al. Creatine kinase isoenzymes in spermatozoa. J Muscle Res Cell Motil 7, 25–34 (1986). https://doi.org/10.1007/BF01756199
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01756199