Different domains of the M-band protein myomesin are involved in myosin binding and M-band targeting.

Myomesin is a 185-kDa protein located in the M-band of striated muscle where it interacts with myosin and titin, possibly connecting thick filaments with the third filament system. By using expression of epitope-tagged myomesin fragments in cultured cardiomyocytes and biochemical binding assays, we could demonstrate that the M-band targeting activity and the myosin-binding site are located in different domains of the molecule. An N-terminal immunoglobulin-like domain is sufficient for targeting to the M-band, but solid-phase overlay assays between individual N-terminal domains and the thick filament protein myosin revealed that the unique head domain contains the myosin-binding site. When expressed in cardiomyocytes, the head domains of rat and chicken myomesin showed species-specific differences in their incorporation pattern. The head domain of rat myomesin localized to a central area within the A-band, whereas the head domain of chicken myomesin was diffusely distributed in the cytoplasm. We therefore conclude that the head domain of myomesin binds to myosin but that this affinity is not sufficient for the restriction of the domain to the M-band in vivo. Instead, the neighboring immunoglobulin-like domain is essential for the precise incorporation of myomesin into the M-band, possibly because of interaction with a yet unknown protein of the sarcomere.

[1]  T. Wallimann,et al.  Myofibrillar interaction of cytosolic creatine kinase (CK) isoenzymes: allocation of N-terminal binding epitope in MM-CK and BB-CK. , 1998, Journal of cell science.

[2]  K. Weber,et al.  Mapping of a myosin-binding domain and a regulatory phosphorylation site in M-protein, a structural protein of the sarcomeric M band. , 1998, Molecular biology of the cell.

[3]  P. F. van der Ven,et al.  Assembly of titin, myomesin and M-protein into the sarcomeric M band in differentiating human skeletal muscle cells in vitro. , 1997, Cell structure and function.

[4]  K Weber,et al.  Molecular structure of the sarcomeric M band: mapping of titin and myosin binding domains in myomesin and the identification of a potential regulatory phosphorylation site in myomesin , 1997, The EMBO journal.

[5]  M. Gautel,et al.  The structure of the sarcomeric M band: localization of defined domains of myomesin, M-protein, and the 250-kD carboxy-terminal region of titin by immunoelectron microscopy , 1996, The Journal of cell biology.

[6]  D. Auerbach,et al.  Tissue-specific Isoforms of Chicken Myomesin Are Generated by Alternative Splicing* , 1996, The Journal of Biological Chemistry.

[7]  J. Perriard,et al.  The intracompartmental sorting of myosin alkali light chain isoproteins reflects the sequence of developmental expression as determined by double epitope-tagging competition. , 1996, Journal of cell science.

[8]  M. Gautel,et al.  A molecular map of titin/connectin elasticity reveals two different mechanisms acting in series , 1996, FEBS letters.

[9]  M. Gautel,et al.  A molecular map of the interactions between titin and myosin-binding protein C. Implications for sarcomeric assembly in familial hypertrophic cardiomyopathy. , 1996, European journal of biochemistry.

[10]  T. Mikawa,et al.  The carboxyl terminus of myosin binding protein C (MyBP-C, C-protein) specifies incorporation into the A-band of striated muscle. , 1996, Journal of cell science.

[11]  Siegfried Labeit,et al.  Titins: Giant Proteins in Charge of Muscle Ultrastructure and Elasticity , 1995, Science.

[12]  D. Fürst,et al.  Purification and biochemical characterization of myomesin, a myosin-binding and titin-binding protein, from bovine skeletal muscle. , 1995, European journal of biochemistry.

[13]  M. Gautel,et al.  The anatomy of a molecular giant: how the sarcomere cytoskeleton is assembled from immunoglobulin superfamily molecules. , 1995, Journal of molecular and cellular cardiology.

[14]  J. Trinick,et al.  Titin and nebulin: protein rulers in muscle? , 1994, Trends in biochemical sciences.

[15]  A. Pastore,et al.  Immunoglobulin‐type domains of titin are stabilized by amino‐terminal extension , 1994, FEBS letters.

[16]  M. Chalfie,et al.  Green fluorescent protein as a marker for gene expression. , 1994, Science.

[17]  F. E. Weber,et al.  The major myosin-binding domain of skeletal muscle MyBP-C (C protein) resides in the COOH-terminal, immunoglobulin C2 motif , 1993, The Journal of cell biology.

[18]  H T van der Voort,et al.  Three-dimensional visualization of multi-channel volume data: the amSFP algorithm. , 1993, Cytometry.

[19]  U. Vinkemeier,et al.  The globular head domain of titin extends into the center of the sarcomeric M band. cDNA cloning, epitope mapping and immunoelectron microscopy of two titin-associated proteins. , 1993, Journal of cell science.

[20]  F. E. Weber,et al.  Complete sequence of human fast-type and slow-type muscle myosin-binding-protein C (MyBP-C). Differential expression, conserved domain structure and chromosome assignment. , 1993, European journal of biochemistry.

[21]  T. Tanaka,et al.  Complete primary structure and tissue expression of chicken pectoralis M-protein. , 1992, The Journal of biological chemistry.

[22]  J. Small,et al.  The cytoskeletal lattice of muscle cells. , 1992, European journal of biochemistry.

[23]  J. Perriard,et al.  Intracompartmental sorting of essential myosin light chains: Molecular dissection and in vivo monitoring by epitope tagging , 1991, Cell.

[24]  J. Trinick.  Elastic filaments and giant proteins in muscle. , 1991, Current opinion in cell biology.

[25]  T Masaki,et al.  Differential distribution of subsets of myofibrillar proteins in cardiac nonstriated and striated myofibrils , 1990, The Journal of cell biology.

[26]  D. Fischman,et al.  Isolation and characterization of a cDNA clone encoding avian skeletal muscle C-protein: an intracellular member of the immunoglobulin superfamily. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[27]  K Weber,et al.  Visualization of the polarity of isolated titin molecules: a single globular head on a long thin rod as the M band anchoring domain? , 1989, The Journal of cell biology.

[28]  K. Weber,et al.  Myogenesis in the mouse embryo: differential onset of expression of myogenic proteins and the involvement of titin in myofibril assembly , 1989, The Journal of cell biology.

[29]  H. Eppenberger,et al.  Fiber type-specific distribution of M-band proteins in chicken muscle. , 1989, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[30]  M. Kozak.  The scanning model for translation: an update , 1989, The Journal of cell biology.

[31]  K. Chien,et al.  Terminally differentiated neonatal rat myocardial cells proliferate and maintain specific differentiated functions following expression of SV40 large T antigen. , 1988, The Journal of biological chemistry.

[32]  J. Perriard,et al.  Intracellular targeting of isoproteins in muscle cytoarchitecture , 1988, The Journal of cell biology.

[33]  M. Price.  Skelemins: cytoskeletal proteins located at the periphery of M-discs in mammalian striated muscle , 1987, The Journal of cell biology.

[34]  E. Kempner,et al.  A physiological role for titin and nebulin in skeletal muscle , 1986, Nature.

[35]  K. Scheidtmann,et al.  Complexes of polyoma virus medium T antigen and cellular proteins. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[36]  H. Eppenberger,et al.  Novel thick filament protein of chicken pectoralis muscle: the 86 kd protein. II. Distribution and localization. , 1985, Journal of molecular biology.

[37]  H. Eppenberger,et al.  Novel thick filament protein of chicken pectoralis muscle: the 86 kd protein. I. Purification and characterization. , 1985, Journal of molecular biology.

[38]  H. Eppenberger,et al.  Myomesin and M-protein: expression of two M-band proteins in pectoral muscle and heart during development , 1985, The Journal of cell biology.

[39]  J. Rose,et al.  A single amino acid substitution in a hydrophobic domain causes temperature-sensitive cell-surface transport of a mutant viral glycoprotein , 1985, Journal of virology.

[40]  C. Lehner,et al.  A new 185,000-dalton skeletal muscle protein detected by monoclonal antibodies , 1984, The Journal of cell biology.

[41]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[42]  C. Kay,et al.  Interaction studies of the 165 000 dalton protein component of the M-line with the S2 subfragment of myosin. , 1978, Biochimica et biophysica acta.

[43]  D. Burgess,et al.  SDS microslab linear gradient polyacrylamide gel electrophoresis. , 1978, Analytical biochemistry.

[44]  R. Craig,et al.  The location of C-protein in rabbit skeletal muscle , 1976, Proceedings of the Royal Society of London. Series B. Biological Sciences.

[45]  O. Takaiti,et al.  M-protein. , 2020, Journal of biochemistry.

[46]  R. Starr,et al.  A new protein of the thick filaments of vertebrate skeletal myofibrils. Extractions, purification and characterization. , 1973, Journal of molecular biology.

[47]  H. Eppenberger,et al.  A protein that binds specifically to the M-line of skeletal muscle is identified as the muscle form of creatine kinase. , 1973, Proceedings of the National Academy of Sciences of the United States of America.

[48]  H. Huxley,et al.  ELECTRON MICROSCOPE STUDIES ON THE STRUCTURE OF NATURAL AND SYNTHETIC PROTEIN FILAMENTS FROM STRIATED MUSCLE. , 1963, Journal of molecular biology.