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Mol Biol Cell. 1996 January; 7(1): 113–127.
PMCID: PMC278617

Modulation of myosin filament organization by C-protein family members.


We have analyzed the interactions between two types of sarcomeric proteins: myosin heavy chain (MyHC) and members of an abundant thick filament-associated protein family (myosin-binding protein; MyBP). Previous work has demonstrated that when MyHC is transiently transfected into mammalian nonmuscle COS cells, the expressed protein forms spindle-shaped structures consisting of bundles of myosin thick filaments. Co-expression of MyHC and MyBP-C or -H modulates the MyHC structures, resulting in dramatically longer cables consisting of myosin and MyBP encircling the nucleus. Immunoelectron microscopy indicates that these cable structures are more uniform in diameter than the spindle structures consisting solely of MyHC, and that the myosin filaments are compacted in the presence of MyBP. Deletion analysis of MyBP-H indicates that cable formation is dependent on the carboxy terminal 24 amino acids. Neither the MyHC spindles nor the MyHC/MyBP cables associate with the endogenous actin cytoskeleton of the COS cell. While there is no apparent co-localization between these structures and the microtubule network, colchicine treatment of the cells promotes the formation of longer assemblages, suggesting that cytoskeletal architecture may physically impede or regulate polymer formation/extension. The data presented here contribute to a greater understanding of the interactions between the MyBP family and MyHC, and provide additional evidence for functional homology between MyBP-C and MyBP-H.

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  • Bähler M, Eppenberger HM, Wallimann T. Novel thick filament protein of chicken pectoralis muscle: the 86 kd protein. I. Purification and characterization. J Mol Biol. 1985 Nov 20;186(2):381–391. [PubMed]
  • Bähler M, Eppenberger HM, Wallimann T. Novel thick filament protein of chicken pectoralis muscle: the 86 kd protein. II. Distribution and localization. J Mol Biol. 1985 Nov 20;186(2):393–401. [PubMed]
  • Bennett P, Craig R, Starr R, Offer G. The ultrastructural location of C-protein, X-protein and H-protein in rabbit muscle. J Muscle Res Cell Motil. 1986 Dec;7(6):550–567. [PubMed]
  • Callaway JE, Bechtel PJ. C-protein from rabbit soleus (red) muscle. Biochem J. 1981 May 1;195(2):463–469. [PubMed]
  • Craig R. Structure of A-segments from frog and rabbit skeletal muscle. J Mol Biol. 1977 Jan 5;109(1):69–81. [PubMed]
  • Davis JS. Interaction of C-protein with pH 8.0 synthetic thick filaments prepared from the myosin of vertebrate skeletal muscle. J Muscle Res Cell Motil. 1988 Apr;9(2):174–183. [PubMed]
  • Dennis JE, Shimizu T, Reinach FC, Fischman DA. Localization of C-protein isoforms in chicken skeletal muscle: ultrastructural detection using monoclonal antibodies. J Cell Biol. 1984 Apr;98(4):1514–1522. [PMC free article] [PubMed]
  • Einheber S, Fischman DA. Isolation and characterization of a cDNA clone encoding avian skeletal muscle C-protein: an intracellular member of the immunoglobulin superfamily. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2157–2161. [PubMed]
  • Feghali R, Leinwand LA. Molecular genetic characterization of a developmentally regulated human perinatal myosin heavy chain. J Cell Biol. 1989 May;108(5):1791–1797. [PMC free article] [PubMed]
  • Hofmann PA, Greaser ML, Moss RL. C-protein limits shortening velocity of rabbit skeletal muscle fibres at low levels of Ca2+ activation. J Physiol. 1991 Aug;439:701–715. [PubMed]
  • Holden HM, Ito M, Hartshorne DJ, Rayment I. X-ray structure determination of telokin, the C-terminal domain of myosin light chain kinase, at 2.8 A resolution. J Mol Biol. 1992 Oct 5;227(3):840–851. [PubMed]
  • Jeacocke SA, England PJ. Phosphorylation of a myofibrillar protein of Mr 150 000 in perfused rat heart, and the tentative indentification of this as C-protein. FEBS Lett. 1980 Dec 15;122(1):129–132. [PubMed]
  • Karsch-Mizrachi I, Feghali R, Shows TB, Leinwand LA. Generation of a full-length human perinatal myosin heavy-chain-encoding cDNA. Gene. 1990 May 14;89(2):289–294. [PubMed]
  • Kaufman RJ, Davies MV, Pathak VK, Hershey JW. The phosphorylation state of eucaryotic initiation factor 2 alters translational efficiency of specific mRNAs. Mol Cell Biol. 1989 Mar;9(3):946–958. [PMC free article] [PubMed]
  • Koretz JF. Effects of C-protein on synthetic myosin filament structure. Biophys J. 1979 Sep;27(3):433–446. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Levy-Mintz P, Kielian M. Mutagenesis of the putative fusion domain of the Semliki Forest virus spike protein. J Virol. 1991 Aug;65(8):4292–4300. [PMC free article] [PubMed]
  • LUFT JH. Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol. 1961 Feb;9:409–414. [PMC free article] [PubMed]
  • Miller JB, Teal SB, Stockdale FE. Evolutionarily conserved sequences of striated muscle myosin heavy chain isoforms. Epitope mapping by cDNA expression. J Biol Chem. 1989 Aug 5;264(22):13122–13130. [PubMed]
  • Moncman CL, Rindt H, Robbins J, Winkelmann DA. Segregated assembly of muscle myosin expressed in nonmuscle cells. Mol Biol Cell. 1993 Oct;4(10):1051–1067. [PMC free article] [PubMed]
  • Moos C. Fluorescence microscope study of the binding of added C protein to skeletal muscle myofibrils. J Cell Biol. 1981 Jul;90(1):25–31. [PMC free article] [PubMed]
  • Moos C, Feng IN. Effect of C-protein on actomyosin ATPase. Biochim Biophys Acta. 1980 Oct 1;632(2):141–149. [PubMed]
  • Moos C, Mason CM, Besterman JM, Feng IN, Dubin JH. The binding of skeletal muscle C-protein to F-actin, and its relation to the interaction of actin with myosin subfragment-1. J Mol Biol. 1978 Oct 5;124(4):571–586. [PubMed]
  • Moos C, Offer G, Starr R, Bennett P. Interaction of C-protein with myosin, myosin rod and light meromyosin. J Mol Biol. 1975 Sep 5;97(1):1–9. [PubMed]
  • Obinata T, Kitani S, Masaki T, Fischman DA. Coexistence of fast-type and slow-type C-proteins in neonatal chicken breast muscle. Dev Biol. 1984 Sep;105(1):253–256. [PubMed]
  • Offer G, Moos C, Starr R. A new protein of the thick filaments of vertebrate skeletal myofibrils. Extractions, purification and characterization. J Mol Biol. 1973 Mar 15;74(4):653–676. [PubMed]
  • Okagaki T, Weber FE, Fischman DA, Vaughan KT, Mikawa T, Reinach FC. The major myosin-binding domain of skeletal muscle MyBP-C (C protein) resides in the COOH-terminal, immunoglobulin C2 motif. J Cell Biol. 1993 Nov;123(3):619–626. [PMC free article] [PubMed]
  • Olson NJ, Pearson RB, Needleman DS, Hurwitz MY, Kemp BE, Means AR. Regulatory and structural motifs of chicken gizzard myosin light chain kinase. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2284–2288. [PubMed]
  • Reinach FC, Masaki T, Fischman DA. Characterization of the C-protein from posterior latissimus dorsi muscle of the adult chicken: heterogeneity within a single sarcomere. J Cell Biol. 1983 Jan;96(1):297–300. [PMC free article] [PubMed]
  • Reinach FC, Masaki T, Shafiq S, Obinata T, Fischman DA. Isoforms of C-protein in adult chicken skeletal muscle: detection with monoclonal antibodies. J Cell Biol. 1982 Oct;95(1):78–84. [PMC free article] [PubMed]
  • Rhee D, Sanger JM, Sanger JW. The premyofibril: evidence for its role in myofibrillogenesis. Cell Motil Cytoskeleton. 1994;28(1):1–24. [PubMed]
  • SABATINI DD, BENSCH K, BARRNETT RJ. Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation. J Cell Biol. 1963 Apr;17:19–58. [PMC free article] [PubMed]
  • Silberstein L, Webster SG, Travis M, Blau HM. Developmental progression of myosin gene expression in cultured muscle cells. Cell. 1986 Sep 26;46(7):1075–1081. [PubMed]
  • Starr R, Almond R, Offer G. Location of C-protein, H-protein and X-protein in rabbit skeletal muscle fibre types. J Muscle Res Cell Motil. 1985 Apr;6(2):227–256. [PubMed]
  • Starr R, Offer G. Polypeptide chains of intermediate molecular weight in myosin preparations. FEBS Lett. 1971 Jun 2;15(1):40–44. [PubMed]
  • Starr R, Offer G. The interaction of C-protein with heavy meromyosin and subfragment-2. Biochem J. 1978 Jun 1;171(3):813–816. [PubMed]
  • Starr R, Offer G. H-protein and X-protein. Two new components of the thick filaments of vertebrate skeletal muscle. J Mol Biol. 1983 Nov 5;170(3):675–698. [PubMed]
  • Takano-Ohmuro H, Goldfine SM, Kojima T, Obinata T, Fischman DA. Size and charge heterogeneity of C-protein isoforms in avian skeletal muscle. Expression of six different isoforms in chicken muscle. J Muscle Res Cell Motil. 1989 Oct;10(5):369–378. [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PubMed]
  • Vaughan KT, Weber FE, Einheber S, Fischman DA. Molecular cloning of chicken myosin-binding protein (MyBP) H (86-kDa protein) reveals extensive homology with MyBP-C (C-protein) with conserved immunoglobulin C2 and fibronectin type III motifs. J Biol Chem. 1993 Feb 15;268(5):3670–3676. [PubMed]
  • Vaughan KT, Weber FE, Fischman DA. cDNA cloning and sequence comparisons of human and chicken muscle C-protein and 86kD protein. Symp Soc Exp Biol. 1992;46:167–177. [PubMed]
  • Vaughan KT, Weber FE, Ried T, Ward DC, Reinach FC, Fischman DA. Human myosin-binding protein H (MyBP-H): complete primary sequence, genomic organization, and chromosomal localization. Genomics. 1993 Apr;16(1):34–40. [PubMed]
  • Vikstrom KL, Rovner AS, Saez CG, Bravo-Zehnder M, Straceski AJ, Leinwand LA. Sarcomeric myosin heavy chain expressed in nonmuscle cells forms thick filaments in the presence of substoichiometric amounts of light chains. Cell Motil Cytoskeleton. 1993;26(3):192–204. [PubMed]
  • Weber FE, Vaughan KT, Reinach FC, Fischman DA. Complete sequence of human fast-type and slow-type muscle myosin-binding-protein C (MyBP-C). Differential expression, conserved domain structure and chromosome assignment. Eur J Biochem. 1993 Sep 1;216(2):661–669. [PubMed]
  • Yamamoto K. Characterization of H-protein, a component of skeletal muscle myofibrils. J Biol Chem. 1984 Jun 10;259(11):7163–7168. [PubMed]
  • Yamamoto K. The binding of skeletal muscle C-protein to regulated actin. FEBS Lett. 1986 Nov 10;208(1):123–127. [PubMed]
  • Yamamoto K. Effect of H-protein on the formation of myosin filaments and light meromyosin paracrystals. J Biochem. 1988 Feb;103(2):274–280. [PubMed]

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