Abstract
Acanthamoeba myosin-I bound to substrates of nitrocellulose or planar lipid membranes on glass moved actin filaments at an average velocity of 0.2 micron/s. This movement required ATP and phosphorylation of the myosin-I heavy chain. We prepared planar lipid membranes on a glass support by passive fusion of lipid vesicles (Brian, A. A., and H. M. McConnell. 1984. Proc. Natl. Acad. Sci. USA. 81:6159-6163) composed of phosphatidylcholine and containing 0-40% phosphatidylserine. The mass of lipid that bound to the glass was the same for membranes of 2 and 20% phosphatidylserine in phosphatidylcholine and was sufficient to form a single bilayer. Myosin-I moved actin filaments on planar membranes of 5-40% but not 0-2% phosphatidylserine. At the low concentrations of phosphatidylserine, actin filaments tended to detach suggesting that less myosin-I was bound. We used the cooperative activation of Acanthamoeba myosin-I ATPase by low concentrations of actin to assess the association of phospholipids with myosin-I. Under conditions where activity depends on the binding of actin to the tail of myosin-I (Albanesi, J. P., H. Fujisaki, and E. D. Korn. 1985. J. Biol. Chem. 260:11174-11179), phospholipid vesicles with 5-40% phosphatidylserine inhibited ATPase activity. The motility and ATPase results demonstrate a specific interaction of the tail of myosin-I with physiological concentrations of phosphatidylserine. This interaction is sufficient to support motility and may provide a mechanism to target myosin-I to biological membranes.
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- Adams R. J., Pollard T. D. Propulsion of organelles isolated from Acanthamoeba along actin filaments by myosin-I. Nature. 1986 Aug 21;322(6081):754–756. doi: 10.1038/322754a0. [DOI] [PubMed] [Google Scholar]
- Albanesi J. P., Fujisaki H., Korn E. D. A kinetic model for the molecular basis of the contractile activity of Acanthamoeba myosins IA and IB. J Biol Chem. 1985 Sep 15;260(20):11174–11179. [PubMed] [Google Scholar]
- Baines I. C., Korn E. D. Localization of myosin IC and myosin II in Acanthamoeba castellanii by indirect immunofluorescence and immunogold electron microscopy. J Cell Biol. 1990 Nov;111(5 Pt 1):1895–1904. doi: 10.1083/jcb.111.5.1895. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berg H. C., Block S. M. A miniature flow cell designed for rapid exchange of media under high-power microscope objectives. J Gen Microbiol. 1984 Nov;130(11):2915–2920. doi: 10.1099/00221287-130-11-2915. [DOI] [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
- Brian A. A., McConnell H. M. Allogeneic stimulation of cytotoxic T cells by supported planar membranes. Proc Natl Acad Sci U S A. 1984 Oct;81(19):6159–6163. doi: 10.1073/pnas.81.19.6159. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brzeska H., Lynch T. J., Korn E. D. Localization of the actin-binding sites of Acanthamoeba myosin IB and effect of limited proteolysis on its actin-activated Mg2+-ATPase activity. J Biol Chem. 1988 Jan 5;263(1):427–435. [PubMed] [Google Scholar]
- Collins K., Sellers J. R., Matsudaira P. Calmodulin dissociation regulates brush border myosin I (110-kD-calmodulin) mechanochemical activity in vitro. J Cell Biol. 1990 Apr;110(4):1137–1147. doi: 10.1083/jcb.110.4.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dabora S. L., Sheetz M. P. The microtubule-dependent formation of a tubulovesicular network with characteristics of the ER from cultured cell extracts. Cell. 1988 Jul 1;54(1):27–35. doi: 10.1016/0092-8674(88)90176-6. [DOI] [PubMed] [Google Scholar]
- De Lozanne A., Spudich J. A. Disruption of the Dictyostelium myosin heavy chain gene by homologous recombination. Science. 1987 May 29;236(4805):1086–1091. doi: 10.1126/science.3576222. [DOI] [PubMed] [Google Scholar]
- Drenckhahn D., Dermietzel R. Organization of the actin filament cytoskeleton in the intestinal brush border: a quantitative and qualitative immunoelectron microscope study. J Cell Biol. 1988 Sep;107(3):1037–1048. doi: 10.1083/jcb.107.3.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fath K. R., Obenauf S. D., Burgess D. R. Cytoskeletal protein and mRNA accumulation during brush border formation in adult chicken enterocytes. Development. 1990 Jun;109(2):449–459. doi: 10.1242/dev.109.2.449. [DOI] [PubMed] [Google Scholar]
- Fukui Y., Lynch T. J., Brzeska H., Korn E. D. Myosin I is located at the leading edges of locomoting Dictyostelium amoebae. Nature. 1989 Sep 28;341(6240):328–331. doi: 10.1038/341328a0. [DOI] [PubMed] [Google Scholar]
- Gadasi H., Korn E. D. Evidence for differential intracellular localization of the Acanthamoeba myosin isoenzymes. Nature. 1980 Jul 31;286(5772):452–456. doi: 10.1038/286452a0. [DOI] [PubMed] [Google Scholar]
- Glenney J. R., Jr, Osborn M., Weber K. The intracellular localization of the microvillus 110K protein, a component considered to be involved in side-on membrane attachment of F-actin. Exp Cell Res. 1982 Mar;138(1):199–205. doi: 10.1016/0014-4827(82)90106-9. [DOI] [PubMed] [Google Scholar]
- Grolig F., Williamson R. E., Parke J., Miller C., Anderton B. H. Myosin and Ca2+-sensitive streaming in the alga Chara: detection of two polypeptides reacting with a monoclonal anti-myosin and their localization in the streaming endoplasm. Eur J Cell Biol. 1988 Oct;47(1):22–31. [PubMed] [Google Scholar]
- Hagen S. J., Kiehart D. P., Kaiser D. A., Pollard T. D. Characterization of monoclonal antibodies to Acanthamoeba myosin-I that cross-react with both myosin-II and low molecular mass nuclear proteins. J Cell Biol. 1986 Dec;103(6 Pt 1):2121–2128. doi: 10.1083/jcb.103.6.2121. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hayden S. M., Wolenski J. S., Mooseker M. S. Binding of brush border myosin I to phospholipid vesicles. J Cell Biol. 1990 Aug;111(2):443–451. doi: 10.1083/jcb.111.2.443. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Horowitz J. A., Hammer J. A., 3rd A new Acanthamoeba myosin heavy chain. Cloning of the gene and immunological identification of the polypeptide. J Biol Chem. 1990 Nov 25;265(33):20646–20652. [PubMed] [Google Scholar]
- Hoshimaru M., Fujio Y., Sobue K., Sugimoto T., Nakanishi S. Immunochemical evidence that myosin I heavy chain-like protein is identical to the 110-kilodalton brush-border protein. J Biochem. 1989 Sep;106(3):455–459. doi: 10.1093/oxfordjournals.jbchem.a122873. [DOI] [PubMed] [Google Scholar]
- Hudspeth A. J. How the ear's works work. Nature. 1989 Oct 5;341(6241):397–404. doi: 10.1038/341397a0. [DOI] [PubMed] [Google Scholar]
- Israelachvili J. N., Marcelja S., Horn R. G. Physical principles of membrane organization. Q Rev Biophys. 1980 May;13(2):121–200. doi: 10.1017/s0033583500001645. [DOI] [PubMed] [Google Scholar]
- Johnson S. M. The effect of charge and cholesterol on the size and thickness of sonicated phospholipid vesicles. Biochim Biophys Acta. 1973 Apr 25;307(1):27–41. doi: 10.1016/0005-2736(73)90022-9. [DOI] [PubMed] [Google Scholar]
- Johnston G. C., Prendergast J. A., Singer R. A. The Saccharomyces cerevisiae MYO2 gene encodes an essential myosin for vectorial transport of vesicles. J Cell Biol. 1991 May;113(3):539–551. doi: 10.1083/jcb.113.3.539. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jung G., Korn E. D., Hammer J. A., 3rd The heavy chain of Acanthamoeba myosin IB is a fusion of myosin-like and non-myosin-like sequences. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6720–6724. doi: 10.1073/pnas.84.19.6720. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jung G., Schmidt C. J., Hammer J. A., 3rd Myosin I heavy-chain genes of Acanthamoeba castellanii: cloning of a second gene and evidence for the existence of a third isoform. Gene. 1989 Oct 30;82(2):269–280. doi: 10.1016/0378-1119(89)90052-8. [DOI] [PubMed] [Google Scholar]
- Kachar B. Direct visualization of organelle movement along actin filaments dissociated from characean algae. Science. 1985 Mar 15;227(4692):1355–1357. doi: 10.1126/science.4038817. [DOI] [PubMed] [Google Scholar]
- Kachar B., Reese T. S. The mechanism of cytoplasmic streaming in characean algal cells: sliding of endoplasmic reticulum along actin filaments. J Cell Biol. 1988 May;106(5):1545–1552. doi: 10.1083/jcb.106.5.1545. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kishino A., Yanagida T. Force measurements by micromanipulation of a single actin filament by glass needles. Nature. 1988 Jul 7;334(6177):74–76. doi: 10.1038/334074a0. [DOI] [PubMed] [Google Scholar]
- Knecht D. A., Loomis W. F. Antisense RNA inactivation of myosin heavy chain gene expression in Dictyostelium discoideum. Science. 1987 May 29;236(4805):1081–1086. doi: 10.1126/science.3576221. [DOI] [PubMed] [Google Scholar]
- Kohno T., Shimmen T. Accelerated sliding of pollen tube organelles along Characeae actin bundles regulated by Ca2+. J Cell Biol. 1988 May;106(5):1539–1543. doi: 10.1083/jcb.106.5.1539. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Korn E. D., Hammer J. A., 3rd Myosin I. Curr Opin Cell Biol. 1990 Feb;2(1):57–61. doi: 10.1016/s0955-0674(05)80031-6. [DOI] [PubMed] [Google Scholar]
- Kron S. J., Spudich J. A. Fluorescent actin filaments move on myosin fixed to a glass surface. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6272–6276. doi: 10.1073/pnas.83.17.6272. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kron S. J., Toyoshima Y. Y., Uyeda T. Q., Spudich J. A. Assays for actin sliding movement over myosin-coated surfaces. Methods Enzymol. 1991;196:399–416. doi: 10.1016/0076-6879(91)96035-p. [DOI] [PubMed] [Google Scholar]
- Lynch T. J., Albanesi J. P., Korn E. D., Robinson E. A., Bowers B., Fujisaki H. ATPase activities and actin-binding properties of subfragments of Acanthamoeba myosin IA. J Biol Chem. 1986 Dec 25;261(36):17156–17162. [PubMed] [Google Scholar]
- Lynch T. J., Brzeska H., Baines I. C., Korn E. D. Purification of myosin I and myosin I heavy chain kinase from Acanthamoeba castellanii. Methods Enzymol. 1991;196:12–23. doi: 10.1016/0076-6879(91)96004-b. [DOI] [PubMed] [Google Scholar]
- McConnell H. M., Watts T. H., Weis R. M., Brian A. A. Supported planar membranes in studies of cell-cell recognition in the immune system. Biochim Biophys Acta. 1986 Jun 12;864(1):95–106. doi: 10.1016/0304-4157(86)90016-x. [DOI] [PubMed] [Google Scholar]
- Mercer J. A., Seperack P. K., Strobel M. C., Copeland N. G., Jenkins N. A. Novel myosin heavy chain encoded by murine dilute coat colour locus. Nature. 1991 Feb 21;349(6311):709–713. doi: 10.1038/349709a0. [DOI] [PubMed] [Google Scholar]
- Miller J. L., Hubbard C. M., Litman B. J., Macdonald T. L. Inhibition of transducin activation and guanosine triphosphatase activity by aluminum ion. J Biol Chem. 1989 Jan 5;264(1):243–250. [PubMed] [Google Scholar]
- Miyata H., Bowers B., Korn E. D. Plasma membrane association of Acanthamoeba myosin I. J Cell Biol. 1989 Oct;109(4 Pt 1):1519–1528. doi: 10.1083/jcb.109.4.1519. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mooseker M. S., Coleman T. R. The 110-kD protein-calmodulin complex of the intestinal microvillus (brush border myosin I) is a mechanoenzyme. J Cell Biol. 1989 Jun;108(6):2395–2400. doi: 10.1083/jcb.108.6.2395. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mooseker M. S., Conzelman K. A., Coleman T. R., Heuser J. E., Sheetz M. P. Characterization of intestinal microvillar membrane disks: detergent-resistant membrane sheets enriched in associated brush border myosin I (110K-calmodulin). J Cell Biol. 1989 Sep;109(3):1153–1161. doi: 10.1083/jcb.109.3.1153. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Poglitsch C. L., Thompson N. L. Interaction of antibodies with Fc receptors in substrate-supported planar membranes measured by total internal reflection fluorescence microscopy. Biochemistry. 1990 Jan 9;29(1):248–254. doi: 10.1021/bi00453a033. [DOI] [PubMed] [Google Scholar]
- Pollard T. D. Assays for myosin. Methods Enzymol. 1982;85(Pt B):123–130. doi: 10.1016/0076-6879(82)85015-5. [DOI] [PubMed] [Google Scholar]
- Pollard T. D., Doberstein S. K., Zot H. G. Myosin-I. Annu Rev Physiol. 1991;53:653–681. doi: 10.1146/annurev.ph.53.030191.003253. [DOI] [PubMed] [Google Scholar]
- Pollard T. D., Korn E. D. Acanthamoeba myosin. II. Interaction with actin and with a new cofactor protein required for actin activation of Mg 2+ adenosine triphosphatase activity. J Biol Chem. 1973 Jul 10;248(13):4691–4697. [PubMed] [Google Scholar]
- Prochniewicz E., Yanagida T. Inhibition of sliding movement of F-actin by crosslinking emphasizes the role of actin structure in the mechanism of motility. J Mol Biol. 1990 Dec 5;216(3):761–772. doi: 10.1016/0022-2836(90)90397-5. [DOI] [PubMed] [Google Scholar]
- Schliwa M., Shimizu T., Vale R. D., Euteneuer U. Nucleotide specificities of anterograde and retrograde organelle transport in Reticulomyxa are indistinguishable. J Cell Biol. 1991 Mar;112(6):1199–1203. doi: 10.1083/jcb.112.6.1199. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schnapp B. J., Reese T. S. Dynein is the motor for retrograde axonal transport of organelles. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1548–1552. doi: 10.1073/pnas.86.5.1548. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schroer T. A., Schnapp B. J., Reese T. S., Sheetz M. P. The role of kinesin and other soluble factors in organelle movement along microtubules. J Cell Biol. 1988 Nov;107(5):1785–1792. doi: 10.1083/jcb.107.5.1785. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schroer T. A., Steuer E. R., Sheetz M. P. Cytoplasmic dynein is a minus end-directed motor for membranous organelles. Cell. 1989 Mar 24;56(6):937–946. doi: 10.1016/0092-8674(89)90627-2. [DOI] [PubMed] [Google Scholar]
- Sellers J. R., Kachar B. Polarity and velocity of sliding filaments: control of direction by actin and of speed by myosin. Science. 1990 Jul 27;249(4967):406–408. doi: 10.1126/science.2377894. [DOI] [PubMed] [Google Scholar]
- Sheetz M. P., Baumrind N. L., Wayne D. B., Pearlman A. L. Concentration of membrane antigens by forward transport and trapping in neuronal growth cones. Cell. 1990 Apr 20;61(2):231–241. doi: 10.1016/0092-8674(90)90804-n. [DOI] [PubMed] [Google Scholar]
- Sheetz M. P., Spudich J. A. Movement of myosin-coated fluorescent beads on actin cables in vitro. Nature. 1983 May 5;303(5912):31–35. doi: 10.1038/303031a0. [DOI] [PubMed] [Google Scholar]
- Small D. M. Phase equilibria and structure of dry and hydrated egg lecithin. J Lipid Res. 1967 Nov;8(6):551–557. [PubMed] [Google Scholar]
- Spudich J. A., Watt S. The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. J Biol Chem. 1971 Aug 10;246(15):4866–4871. [PubMed] [Google Scholar]
- Titus M. A., Warrick H. M., Spudich J. A. Multiple actin-based motor genes in Dictyostelium. Cell Regul. 1989 Nov;1(1):55–63. doi: 10.1091/mbc.1.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Toyoshima Y. Y., Kron S. J., McNally E. M., Niebling K. R., Toyoshima C., Spudich J. A. Myosin subfragment-1 is sufficient to move actin filaments in vitro. Nature. 1987 Aug 6;328(6130):536–539. doi: 10.1038/328536a0. [DOI] [PubMed] [Google Scholar]
- Ulsamer A. G., Wright P. L., Wetzel M. G., Korn E. D. Plasma and phagosome membranes of Acanthamoeba castellanii. J Cell Biol. 1971 Oct;51(1):193–215. doi: 10.1083/jcb.51.1.193. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Umemoto S., Sellers J. R. Characterization of in vitro motility assays using smooth muscle and cytoplasmic myosins. J Biol Chem. 1990 Sep 5;265(25):14864–14869. [PubMed] [Google Scholar]
- Vale R. D., Hotani H. Formation of membrane networks in vitro by kinesin-driven microtubule movement. J Cell Biol. 1988 Dec;107(6 Pt 1):2233–2241. doi: 10.1083/jcb.107.6.2233. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Warshaw D. M., Desrosiers J. M., Work S. S., Trybus K. M. Smooth muscle myosin cross-bridge interactions modulate actin filament sliding velocity in vitro. J Cell Biol. 1990 Aug;111(2):453–463. doi: 10.1083/jcb.111.2.453. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yamada A., Ishii N., Takahashi K. Direction and speed of actin filaments moving along thick filaments isolated from molluscan smooth muscle. J Biochem. 1990 Sep;108(3):341–343. doi: 10.1093/oxfordjournals.jbchem.a123203. [DOI] [PubMed] [Google Scholar]