Summary
We have determined by protein chemistry methods the amino acid sequence of light chain 2 fromAcanthamoeba castellanii myosin-II (ALC2). This is the first reported sequence for any protozoan myosin light chain. ALC2 consists of 154 amino acid residues, including a single residue of His and two residues each of Pro and Tyr, and lacks Cys and Trp. The N-terminus is blocked, and if an N-terminal acetyl group is assumed, ALC2 has a calculated molecular weight of 17657. ALC2 is an acidic protein, with a calculated net charge of — 7 at pH 7. The sequence of ALC2 is most similar to those of the calmodulins (identity approximately 35%, followed by myosin regulatory light chains. ALC2 appears to lack the potential N-terminal phosphorylation site and single Ca2+-binding site in region I which are characteristic of most myosin regulatory light chains. Instead, ALC2, unlike any other myosin light chain characterized to date, may have a functional Ca2+-binding site only in region II, suggesting a novel role of ALC2 in the Ca2+ regulation of the activity ofAcanthamoeba myosin-II.
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References
Baba, M. L., Goodman, M., Berger-Cohn, J., Demaille, J. G. &Matsuda, G. (1984) The early adaptive evolution of calmodulin.Molec. Biol. Evol. 1, 442–55.
Babu, Y. S., Sack, J. S., Greenhough, T. J., Bugg, G. E., Means, A. R. &Cook, W. J. (1985) Three-dimensional structure of calmodulin.Nature 315, 37–40.
Babu, Y. S., Bugg, C. E. &Cook, W. J. (1988) Structure of calmodulin refined at 2.2 Å resolution.J. Mol. Biol. 204, 191–204.
Chisolm, R. L., Rushforth, A. M., Pollenz, R. S., Kuczmarski, E. R. &Tafuri, S. R. (1988)Dictiostelium discoideum myosin: isolation and characterization of cDNAs encoding the essential light chain.Mol. Cell. Biol. 8, 794–801.
Collins, J. H. (1974) Homology of myosin light chains, troponin C and parvalbumins deduced from comparison of their amino acid sequences.Biochem. Biophys. Res. Commun. 58, 301–8.
Collins, J. H. (1976) Structure and evolution of troponin C and related proteins.Soc. Exp. Biol. Symp. 30, 303–34.
Collins, J. H. (1991) Myosin light chains and troponin C: structural and evolutionary relationships revealed by amino acid sequence comparisons.J. Muscle Res. Cell Motil. 12, 3–25.
Collins, J. H. &Korn, E. D. (1980) Actin activation of Ca2+- sensitive Mg2+-ATPase activity ofAcanthamoeba myosin II is enhanced by dephosphorylation of its heavy chains.J. Biol. Chem. 255, 8011–14.
Collins, J. H. &Korn, E. D. (1981) Purification and characterization of actin-activateable, Ca-sensitive myosin-II fromAcanthamoeba.J. Biol. Chem. 256, 2586–95.
Collins, J. K., Potter, J. D., Horn, M. J., Wilshire, G. &Jackman, N. (1973) The amino acid sequence of rabbit skeletal muscle troponin C: gene replication and homology with calcium-binding proteins from carp and hake muscle.FEBS Lett. 36, 268–72.
Collins, J. H., Jakes, R., Kendrick-Jones, J., Leszyk, J., Barouch, W., Theibert, J. L., Spiegel, J. &Szent-Gyorgyi, A. G. (1986) Amino acid sequence of myosin essential light chain from the scallopAquipecten irradians.Biochemistry 25, 7651–6.
Coté, G. P., Collins, J. H., &Korn, E. D. (1981) Identification of three phosphorylation sites on each heavy chain ofAcanthamoeba myosin II.J. Biol. Chem. 256, 12811–16.
Goodwin, E. B., Szent-Gyorgyi, A. G., &Leinwand, L. (1987) Cloning and characterization of the scallop essential and regulatory myosin light chain cDNAs.J. Biol. Chem. 262, 11052–6.
Griffith, L. M., Downs, S. M. &Spudich, J. A. (1987) Myosin light chain kinase and myosin light chain phosphatase fromDictyostelium:effects of reversible phosphorylation on myosin structure and function.J. Cell Biol. 104, 1309–23.
Hammer, J. A. III, Bowers, B., Paterson, B. M. &Korn, E. D. (1987) Complete nucleotide sequence and deduced polypeptide sequence of a non-muscle myosin heavy chain gene fromAcanthamoeba: evidence of a hinge in the rodlike tail.J. Cell Biol. 105, 913–25.
Heinrickson, R. L. &Meredith, S. C. (1984) Amino acid analysis by reverse-phase high-performance liquid chromatography: precolumn derivatization with phenylisothiocyanate.Anal. Biochem. 136, 65–74.
Herzberg, O. &James, M. N. G. (1985) Structure of the calcium regulatory muscle protein troponin-C at 2.8 Å resolution.Nature 313, 653–9.
Herzberg, O. &James, M. N. G. (1988) Refined crystal structure of troponin C from turkey skeletal muscle at 2.0 Å resolution.J. Mol. Biol. 203, 761–79.
Kendrick-Jones, J., Lehman, W. &Szent-Gyorgyi, A. G. (1970) Regulation in molluscan muscles.J. Molec. Biol. 54, 313–26.
Klee, C. B. (1977) Conformational transition accompanying the binding of Ca2+ to the protein activator of 3′, 5′-cyclic adenosine monophosphate phosphodiesterase.Biochemistry 16, 1017–24.
Kobayashi, T., Takagi, T., Konishi, K., Hamada, Y., Kawaguchi, M. &Kohama, K. (1988) Amino acid sequence of the calcium-binding light chain of myosin from the lower eukaryotePhysarum polycephalum.J. Biol. Chem. 263, 305–13.
Kohama, K. (1989) Ca-binding light chain of Ca-inhibitory myosin fromPhysarum polycephalum. InCalcium Signal and Cell Response (edited byYagi, K. &Miyazaki, T.) pp. 95–105 Tokyo:Japan Scientific Society Press, and Berlin:Springer.
Korn E. D. &Hammer, J. A. III (1988) Myosins of non-muscle cells.Ann. Rev. Biophys. Biophys. Chem. 17, 23–45.
Korn, E. D., Atkinson, M. A. L., Brzeska, H., Hammer, J. A. III, Jung, G. &Lynch, T. J. (1988) Structure-function studies onAcanthamoeba myosins IA, IB and II.J. Cell. Biochem. 36, 37–50.
Kwon, H., Goodwin, E. B., Nyitray, L., Berliner, E., O'Neall-Hennessey, E., Melandri, F. D. &Szent-Gyorgyi, A. G. (1990) Isolation of the regulatory domain of scallop myosin: role of the essential light chain in calcium binding.Proc. Natl Acad. Sci. (USA) 87, 4771–5.
Leavis, P. C. &Gergely, J. (1984) Thin filament proteins and thin filament-linked regulation of vertebrate muscle contraction.CRC Crit. Rev. Biochem. 16, 235–305.
Malencik, D. A. &Anderson, S. R. (1988) Peptide cross-linking to calmodulin: attachment of [Tyr8] substance P.Biochemistry 27, 1941–9.
Marshak, D. R., Clarke, M., Roberts, D. M. &Watterson, D. M. (1984) Structural and functional properties of calmodulin from the eukaryotic microorganismDictyostelium discoideum.Biochemistry 23, 2891–9.
Maruta, H. &Korn, E. D. (1977)Acanthamoeba myosin II.J. Biol. Chem. 252, 6501–9.
Satyshur, K. A., Rao, S. T., Pyzalska, D., Drendel., W., Greaser, M. &Sundaralingam, M. (1988) Refined structure of chicken skeletal muscle troponin C in the two-calcium state at 2-A resolution.J. Biol. Chem. 263, 1628–47.
Sinard, J. H. &Pollard, T. D. (1989) The effect of heavy chain phosphorylation and solution conditions on the assembly ofAcanthamoeba myosin-II.J. Cell Biol. 109, 1529–35.
Sundaralingam, M., Bergstrom, R., Strasburg, G., Rao, S. T., Roychowdhury, P., Greaser, M. &Wang, B. C. (1985) Molecular structure of troponin C from chicken skeletal muscle at 3-angstrom resolution.Science 227, 945–8.
Tafuri, S. R., Rushforth, A. M., Kuczmarski, E. R. &Chisolm, R. L. (1989)Dictiostelium discoideum myosin: isolation and characterization of cDNAs encoding the regulatory light chain.Mol. Cell. Biol. 9, 3073–80.
Warrick, H. M., Delozanne, A., Leinwand, L. A. &Spudich, J. A. (1986) Conserved protein domains in a myosin heavy chain gene fromDictyostelium discoideum.Proc. Natl Acad. Sci. (USA) 83, 9433–7.
Zot, A. S. &Potter, J. D. (1987) Structural aspects of troponintropomyosin regulation of skeletal muscle contraction.Ann. Rev. Biophys. Biophys. Chem. 16, 535–59.
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Kobayashi, T., Zot, H.G., Pollard, T.D. et al. Functional implications of the unusual amino acid sequence of the regulatory light chain ofAcanthamoeba castellanii myosin-II. J Muscle Res Cell Motil 12, 553–559 (1991). https://doi.org/10.1007/BF01738443
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DOI: https://doi.org/10.1007/BF01738443