Skip to main content
PMC home page
Biophysical Journal logoLink to Biophysical Journal
. 1995 Apr;68(4 Suppl):112S–119S.

Function of the N terminus of the myosin essential light chain of vertebrate striated muscle.

H L Sweeney 1
PMCID: PMC1281889  PMID: 7787052

Abstract

All but one (LC3-f; a fast skeletal muscle isoform) of the essential light chain isoforms of myosin (ELC) that are expressed in vertebrate striated muscles have an extended N terminus that is found neither in invertebrate ELCs nor in the majority of vertebrate smooth and nonmuscle myosin ELCs. Studies with permeabilized skeletal muscle fibers and in vitro motility assays have demonstrated that the presence of the ELC isoform lacking the N-terminal extension (LC3-f) is correlated with an increased maximal velocity of filament sliding. To examine further this modulatory role of the ELCs, a procedure was developed for the exchange of ELCs that is based on a technique for the removal of regulatory light chains from permeabilized muscle fibers. Different isoforms of the ELCs and mutant ELCs were exchanged into permeabilized skeletal muscle fibers from rabbit psoas muscle. The role of the ELCs of myosin in altering the shortening Vmax of striated muscle was confirmed. Additionally, experiments with mutant ELCs in which lysines at the extreme N terminus were replaced with alanines, demonstrated an increased shortening Vmax that coincided with removal of the positive charges contributed by the lysines. This suggests that charge interactions (i.e., salt bridges) between the N terminus of the ELC and negatively charged amino acids on the surface of actin cause a slowing of filament sliding. Whether this role in altering shortening velocity is the primary function of the extended N terminus of the ELC or whether it is merely a consequence of providing a tether between the thick and thin filaments is discussed.

Full text

PDF
118s

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bottinelli R., Betto R., Schiaffino S., Reggiani C. Maximum shortening velocity and coexistence of myosin heavy chain isoforms in single skinned fast fibres of rat skeletal muscle. J Muscle Res Cell Motil. 1994 Aug;15(4):413–419. doi: 10.1007/BF00122115. [DOI] [PubMed] [Google Scholar]
  2. Butler-Browne G. S., Whalen R. G. Myosin isozyme transitions occurring during the postnatal development of the rat soleus muscle. Dev Biol. 1984 Apr;102(2):324–334. doi: 10.1016/0012-1606(84)90197-0. [DOI] [PubMed] [Google Scholar]
  3. Bárány K., Bárány M., Gillis J. M., Kushmerick M. J. Phosphorylation-dephosphorylation of the 18,000-dalton light chain of myosin during the contraction-relaxation cycle of frog muscle. J Biol Chem. 1979 May 10;254(9):3617–3623. [PubMed] [Google Scholar]
  4. Chaussepied P., Kasprzak A. A. Isolation and characterization of the G-actin-myosin head complex. Nature. 1989 Dec 21;342(6252):950–953. doi: 10.1038/342950a0. [DOI] [PubMed] [Google Scholar]
  5. Collins J. H. Myosin light chains and troponin C: structural and evolutionary relationships revealed by amino acid sequence comparisons. J Muscle Res Cell Motil. 1991 Feb;12(1):3–25. doi: 10.1007/BF01781170. [DOI] [PubMed] [Google Scholar]
  6. Craig R., Padrón R., Kendrick-Jones J. Structural changes accompanying phosphorylation of tarantula muscle myosin filaments. J Cell Biol. 1987 Sep;105(3):1319–1327. doi: 10.1083/jcb.105.3.1319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eastwood A. B., Wood D. S., Bock K. L., Sorenson M. M. Chemically skinned mammalian skeletal muscle. I. The structure of skinned rabbit psoas. Tissue Cell. 1979;11(3):553–566. doi: 10.1016/0040-8166(79)90062-4. [DOI] [PubMed] [Google Scholar]
  8. Edman K. A. The velocity of unloaded shortening and its relation to sarcomere length and isometric force in vertebrate muscle fibres. J Physiol. 1979 Jun;291:143–159. doi: 10.1113/jphysiol.1979.sp012804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Frank G., Weeds A. G. The amino-acid sequence of the alkali light chains of rabbit skeletal-muscle myosin. Eur J Biochem. 1974 May 15;44(2):317–334. doi: 10.1111/j.1432-1033.1974.tb03489.x. [DOI] [PubMed] [Google Scholar]
  10. Gauthier G. F., Lowey S., Hobbs A. W. Fast and slow myosin in developing muscle fibres. Nature. 1978 Jul 6;274(5666):25–29. doi: 10.1038/274025a0. [DOI] [PubMed] [Google Scholar]
  11. Gazith J., Himmelfarb S., Harrington W. F. Studies on the subunit structure of myosin. J Biol Chem. 1970 Jan 10;245(1):15–22. [PubMed] [Google Scholar]
  12. Giulian G. G., Moss R. L., Greaser M. Improved methodology for analysis and quantitation of proteins on one-dimensional silver-stained slab gels. Anal Biochem. 1983 Mar;129(2):277–287. doi: 10.1016/0003-2697(83)90551-1. [DOI] [PubMed] [Google Scholar]
  13. Greaser M. L., Moss R. L., Reiser P. J. Variations in contractile properties of rabbit single muscle fibres in relation to troponin T isoforms and myosin light chains. J Physiol. 1988 Dec;406:85–98. doi: 10.1113/jphysiol.1988.sp017370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Güth K., Wojciechowski R. Perfusion cuvette for the simultaneous measurement of mechanical, optical and energetic parameters of skinned muscle fibres. Pflugers Arch. 1986 Nov;407(5):552–557. doi: 10.1007/BF00657515. [DOI] [PubMed] [Google Scholar]
  15. Hailstones D. L., Gunning P. W. Characterization of human myosin light chains 1sa and 3nm: implications for isoform evolution and function. Mol Cell Biol. 1990 Mar;10(3):1095–1104. doi: 10.1128/mcb.10.3.1095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Henry G. D., Winstanley M. A., Dalgarno D. C., Scott G. M., Levine B. A., Trayer I. P. Characterization of the actin-binding site on the alkali light chain of myosin. Biochim Biophys Acta. 1985 Aug 23;830(3):233–243. doi: 10.1016/0167-4838(85)90279-1. [DOI] [PubMed] [Google Scholar]
  17. Hoh J. Y., McGrath P. A., White R. I. Electrophoretic analysis of multiple forms of myosin in fast-twitch and slow-twitch muscles of the chick. Biochem J. 1976 Jul 1;157(1):87–95. doi: 10.1042/bj1570087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Johara M., Toyoshima Y. Y., Ishijima A., Kojima H., Yanagida T., Sutoh K. Charge-reversion mutagenesis of Dictyostelium actin to map the surface recognized by myosin during ATP-driven sliding motion. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2127–2131. doi: 10.1073/pnas.90.6.2127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Julian F. J., Moss R. L., Waller G. S. Mechanical properties and myosin light chain composition of skinned muscle fibres from adult and new-born rabbits. J Physiol. 1981 Feb;311:201–218. doi: 10.1113/jphysiol.1981.sp013581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kamm K. E., Stull J. T. The function of myosin and myosin light chain kinase phosphorylation in smooth muscle. Annu Rev Pharmacol Toxicol. 1985;25:593–620. doi: 10.1146/annurev.pa.25.040185.003113. [DOI] [PubMed] [Google Scholar]
  21. LaFramboise W. A., Daood M. J., Guthrie R. D., Moretti P., Schiaffino S., Ontell M. Electrophoretic separation and immunological identification of type 2X myosin heavy chain in rat skeletal muscle. Biochim Biophys Acta. 1990 Jul 20;1035(1):109–112. doi: 10.1016/0304-4165(90)90181-u. [DOI] [PubMed] [Google Scholar]
  22. Labbé J. P., Audemard E., Bertrand R., Kassab R. Specific interactions of the alkali light chain 1 in skeletal myosin heads probed by chemical cross-linking. Biochemistry. 1986 Dec 16;25(25):8325–8330. doi: 10.1021/bi00373a029. [DOI] [PubMed] [Google Scholar]
  23. Levine R. J., Chantler P. D., Kensler R. W., Woodhead J. L. Effects of phosphorylation by myosin light chain kinase on the structure of Limulus thick filaments. J Cell Biol. 1991 May;113(3):563–572. doi: 10.1083/jcb.113.3.563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lheureux K., Forné T., Chaussepied P. Interaction and polymerization of the G-actin-myosin head complex: effect of DNase I. Biochemistry. 1993 Sep 28;32(38):10005–10014. doi: 10.1021/bi00089a016. [DOI] [PubMed] [Google Scholar]
  25. Lowey S., Waller G. S., Trybus K. M. Skeletal muscle myosin light chains are essential for physiological speeds of shortening. Nature. 1993 Sep 30;365(6445):454–456. doi: 10.1038/365454a0. [DOI] [PubMed] [Google Scholar]
  26. Mabuchi K., Szvetko D., Pintér K., Sréter F. A. Type IIB to IIA fiber transformation in intermittently stimulated rabbit muscles. Am J Physiol. 1982 May;242(5):C373–C381. doi: 10.1152/ajpcell.1982.242.5.C373. [DOI] [PubMed] [Google Scholar]
  27. Margossian S. S., Bhan A. K., Slayter H. S. Role of the regulatory light chains in skeletal muscle actomyosin ATPase and in minifilament formation. J Biol Chem. 1983 Nov 10;258(21):13359–13369. [PubMed] [Google Scholar]
  28. Merril C. R., Goldman D. Quantitative two-dimensional protein electrophoresis for studies of inborn errors of metabolism. Clin Chem. 1982 Apr;28(4 Pt 2):1015–1020. [PubMed] [Google Scholar]
  29. Metzger J. M., Greaser M. L., Moss R. L. Variations in cross-bridge attachment rate and tension with phosphorylation of myosin in mammalian skinned skeletal muscle fibers. Implications for twitch potentiation in intact muscle. J Gen Physiol. 1989 May;93(5):855–883. doi: 10.1085/jgp.93.5.855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Miedema K., Harhangi H., Mentzel S., Wilbrink M., Akhmanova A., Hooiveld M., Bindels P., Hennig W. Interspecific sequence comparison of the muscle-myosin heavy-chain genes from Drosophila hydei and Drosophila melanogaster. J Mol Evol. 1994 Oct;39(4):357–368. doi: 10.1007/BF00160268. [DOI] [PubMed] [Google Scholar]
  31. Milligan R. A., Whittaker M., Safer D. Molecular structure of F-actin and location of surface binding sites. Nature. 1990 Nov 15;348(6298):217–221. doi: 10.1038/348217a0. [DOI] [PubMed] [Google Scholar]
  32. Pastra-Landis S. C., Lowey S. Myosin subunit interactions. Properties of the 19,000-dalton light chain-deficient myosin. J Biol Chem. 1986 Nov 5;261(31):14811–14816. [PubMed] [Google Scholar]
  33. Periasamy M., Strehler E. E., Garfinkel L. I., Gubits R. M., Ruiz-Opazo N., Nadal-Ginard B. Fast skeletal muscle myosin light chains 1 and 3 are produced from a single gene by a combined process of differential RNA transcription and splicing. J Biol Chem. 1984 Nov 10;259(21):13595–13604. [PubMed] [Google Scholar]
  34. Prince H. P., Trayer H. R., Henry G. D., Trayer I. P., Dalgarno D. C., Levine B. A., Cary P. D., Turner C. Proton nuclear-magnetic-resonance spectroscopy of myosin subfragment 1 isoenzymes. Eur J Biochem. 1981 Dec;121(1):213–219. doi: 10.1111/j.1432-1033.1981.tb06451.x. [DOI] [PubMed] [Google Scholar]
  35. Putkey J. A., Liu W., Sweeney H. L. Function of the N-terminal calcium-binding sites in cardiac/slow troponin C assessed in fast skeletal muscle fibers. J Biol Chem. 1991 Aug 15;266(23):14881–14884. [PubMed] [Google Scholar]
  36. Rayment I., Rypniewski W. R., Schmidt-Bäse K., Smith R., Tomchick D. R., Benning M. M., Winkelmann D. A., Wesenberg G., Holden H. M. Three-dimensional structure of myosin subfragment-1: a molecular motor. Science. 1993 Jul 2;261(5117):50–58. doi: 10.1126/science.8316857. [DOI] [PubMed] [Google Scholar]
  37. Reiser P. J., Moss R. L., Giulian G. G., Greaser M. L. Shortening velocity in single fibers from adult rabbit soleus muscles is correlated with myosin heavy chain composition. J Biol Chem. 1985 Aug 5;260(16):9077–9080. [PubMed] [Google Scholar]
  38. Sivaramakrishnan M., Burke M. The free heavy chain of vertebrate skeletal myosin subfragment 1 shows full enzymatic activity. J Biol Chem. 1982 Jan 25;257(2):1102–1105. [PubMed] [Google Scholar]
  39. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  40. Sutoh K., Ando M., Sutoh K., Toyoshima Y. Y. Site-directed mutations of Dictyostelium actin: disruption of a negative charge cluster at the N terminus. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7711–7714. doi: 10.1073/pnas.88.17.7711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Sutoh K. Identification of myosin-binding sites on the actin sequence. Biochemistry. 1982 Jul 20;21(15):3654–3661. doi: 10.1021/bi00258a020. [DOI] [PubMed] [Google Scholar]
  42. Sweeney H. L., Bowman B. F., Stull J. T. Myosin light chain phosphorylation in vertebrate striated muscle: regulation and function. Am J Physiol. 1993 May;264(5 Pt 1):C1085–C1095. doi: 10.1152/ajpcell.1993.264.5.C1085. [DOI] [PubMed] [Google Scholar]
  43. Sweeney H. L., Corteselli S. A., Kushmerick M. J. Measurements on permeabilized skeletal muscle fibers during continuous activation. Am J Physiol. 1987 May;252(5 Pt 1):C575–C580. doi: 10.1152/ajpcell.1987.252.5.C575. [DOI] [PubMed] [Google Scholar]
  44. Sweeney H. L., Kushmerick M. J., Mabuchi K., Sréter F. A., Gergely J. Myosin alkali light chain and heavy chain variations correlate with altered shortening velocity of isolated skeletal muscle fibers. J Biol Chem. 1988 Jun 25;263(18):9034–9039. [PubMed] [Google Scholar]
  45. Sweeney H. L., Kushmerick M. J. Myosin phosphorylation in permeabilized rabbit psoas fibers. Am J Physiol. 1985 Sep;249(3 Pt 1):C362–C365. doi: 10.1152/ajpcell.1985.249.3.C362. [DOI] [PubMed] [Google Scholar]
  46. Sweeney H. L., Stull J. T. Alteration of cross-bridge kinetics by myosin light chain phosphorylation in rabbit skeletal muscle: implications for regulation of actin-myosin interaction. Proc Natl Acad Sci U S A. 1990 Jan;87(1):414–418. doi: 10.1073/pnas.87.1.414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Sweeney H. L., Stull J. T. Phosphorylation of myosin in permeabilized mammalian cardiac and skeletal muscle cells. Am J Physiol. 1986 Apr;250(4 Pt 1):C657–C660. doi: 10.1152/ajpcell.1986.250.4.C657. [DOI] [PubMed] [Google Scholar]
  48. Sweeney H. L., Yang Z., Zhi G., Stull J. T., Trybus K. M. Charge replacement near the phosphorylatable serine of the myosin regulatory light chain mimics aspects of phosphorylation. Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1490–1494. doi: 10.1073/pnas.91.4.1490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Trayer I. P., Trayer H. R., Levine B. A. Evidence that the N-terminal region of A1-light chain of myosin interacts directly with the C-terminal region of actin. A proton magnetic resonance study. Eur J Biochem. 1987 Apr 1;164(1):259–266. doi: 10.1111/j.1432-1033.1987.tb11019.x. [DOI] [PubMed] [Google Scholar]
  50. Wade R., Feldman D., Gunning P., Kedes L. Sequence and expression of human myosin alkali light chain isoforms. Mol Cell Biochem. 1989 Jun 1;87(2):119–136. doi: 10.1007/BF00219255. [DOI] [PubMed] [Google Scholar]
  51. Wagner P. D., Giniger E. Hydrolysis of ATP and reversible binding to F-actin by myosin heavy chains free of all light chains. Nature. 1981 Aug 6;292(5823):560–562. doi: 10.1038/292560a0. [DOI] [PubMed] [Google Scholar]
  52. Wagner P. D. Preparation and fractionation of myosin light chains and exchange of the essential light chains. Methods Enzymol. 1982;85(Pt B):72–81. doi: 10.1016/0076-6879(82)85010-6. [DOI] [PubMed] [Google Scholar]
  53. Wagner P. D., Weeds A. G. Studies on the role of myosin alkali light chains. Recombination and hybridization of light chains and heavy chains in subfragment-1 preparations. J Mol Biol. 1977 Jan 25;109(3):455–470. doi: 10.1016/s0022-2836(77)80023-5. [DOI] [PubMed] [Google Scholar]
  54. Weeds A. G. Light chains from slow-twitch muscle myosin. Eur J Biochem. 1976 Jun 15;66(1):157–173. doi: 10.1111/j.1432-1033.1976.tb10436.x. [DOI] [PubMed] [Google Scholar]
  55. Weeds A. G. Light chains of myosin. Nature. 1969 Sep 27;223(5213):1362–1364. doi: 10.1038/2231362a0. [DOI] [PubMed] [Google Scholar]
  56. Weeds A. G., Lowey S. Substructure of the myosin molecule. II. The light chains of myosin. J Mol Biol. 1971 Nov 14;61(3):701–725. doi: 10.1016/0022-2836(71)90074-x. [DOI] [PubMed] [Google Scholar]
  57. Whalen R. G., Sell S. M., Butler-Browne G. S., Schwartz K., Bouveret P., Pinset-Härstöm I. Three myosin heavy-chain isozymes appear sequentially in rat muscle development. Nature. 1981 Aug 27;292(5826):805–809. doi: 10.1038/292805a0. [DOI] [PubMed] [Google Scholar]
  58. Winstanley M. A., Trayer I. P. Thrombic digestion of myosin alkali light chains [proceedings]. Biochem Soc Trans. 1979 Aug;7(4):703–704. doi: 10.1042/bst0070703. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES