Cooperation between the two myosin heads interacting with actin in presence of ADP in myofibrils

Zusammenfassung Alkylierung von 2 Thiolgruppen pro Myosin mit NEM bei 0°C in Gegenwart einer Diophosphatkette inaktiviert die K-ATPase vollständig.In Myofibrillen werden diese Thiolgruppen durch Rigor-interaction beider Myosinköpfchen mit Actin vor Alkylierung geschützt. In Gegenwart von Mg-ADP tritt eine spezifische vom Rigor verschiedene interaction zwischen Myosin und Actin auf. Man muss annehemen, dass dabei nur 1 Myosinköpfchen ans Actin bindet und dass das andere Köpfchen ein ADP gebunden hat.     

The fossil vertebrates from East Greenland and their zoological importance

Zusammenfassung Kurzer Bericht über die dänischen Expeditionen nach Ostgrönland (Lauge Koch) 1929–1957. Die zoologische Bedeutung des fossilen Vertebratenmaterials (über 10000 Exemplare) wird angedeutet. Neuere Ergebnisse über die Evolution der Tetrapodenzunge werden besonders im Zusammenhang mit Grönlandmaterial besprochen. Damit wird die Ansicht bestätigt, dass sich die Urodelen selbständig aus porolepiformen Crossopterygiern entwickelt haben, während Anuren, Sauropsiden und Säugetiere von Osteolepiformiern abzuleiten sind. Die Auswertung des umfangreichen Fossilmaterials ergibt eine diphyletische Herkunft der Tetrapoden.     

Metal binding to myosin and to myosin DTNB-light chain

Summary The effects of various divalent cations, Ca²⁺, Mg²⁺ and Mn²⁺ on the intrinsic fluorescence of heavy meromyosin (HMM) and myosin 5,5-dithio-bis-(2-nitrobenzoate) DTNB-light chain of rabbit striated muscle, are compared. At pH 6.4, the fluorescence change induced by the metal ions is present only in the isolated light chain and disappears in HMM, thus indicating an interaction between the heavy and light chains with respect to the binding of the metal ions. Whereas Mg²⁺ binds more strongly than Ca²⁺ to myosin, this order is reversed in the case of the DTNB-light chain.     

Electron-microscopic mapping of the hinge region of myosin

Summary The trypsin-sensitive sites in the labile hinge region of the myosin molecule are located with heightened accuracy (±2 nm) by electron microscopy as lying at 70, 85, 95, and 103 nm from the C-terminus of the rod section of the molecule.I thank Dr.Michael Young for valuable advice and encouragement, Drs.Jerome Gross andRomaine R. Bruns for use of their electron microscope, and Mrs.Muriel H. Blanchard for excellent technical assistance. This work was supported by the American Heart Association, The John A. Hartford Foundation, Inc., and the National Institutes of Health.     

Effects of cytochalasin D on the actin cytoskeleton: association of neoformed actin aggregates with proteins involved in signaling and endocytosis

Cytochalasin D (CD) has been extensively used for assessing the role of the actin cytoskeleton in different biological processes. However, effects of CD have not always been consistent and CD-treated cells have been found to contain irregular spots of F-actin. By transfecting MCF-7 cells with an actin-enhanced yellow fluorescent protein fusion protein we show that, in vivo, CD induces actin aggregation de novo, while simultaneously depolymerizing preexisting actin cytoskeletal components. We also show that CD-induced actin aggregates bind the F-actin-selective drug phalloidin and associate with proteins involved in cell signaling as well as with receptors and endosomal markers (active MAP kinases, paxillin, erbB2, transferrin, Rab-5), but not with clathrin, protein kinase A, protein tyrosine phosphatase 1B, or tubulin. Thus, CD induces new sites of actin aggregation that selectively associate with several important regulatory proteins. Failure of CD to interupt a biological process may therefore not prove that the process is independent of actin aggregation.     

Regulation of class V myosin

Class V myosin (myosin-5) is a molecular motor that functions as an organelle transporter. The activation of myosin-5′s motor function has long been known to be associated with a transition from the folded conformation in the off-state to the extended conformation in the on-state, but only recently have we begun to understand the underlying mechanism. The globular tail domain (GTD) of myosin-5 has been identified as the inhibitory domain and has recently been shown to function as a dimer in regulating the motor function. The folded off-state of myosin-5 is stabilized by multiple intramolecular interactions, including head–GTD interactions, GTD–GTD interactions, and interactions between the GTD and the C-terminus of the first coiled-coil segment. Any cellular factor that affects these intramolecular interactions and thus the stability of the folded conformation of myosin-5 would be expected to regulate myosin-5 motor function. Both the adaptor proteins of myosin-5 and Ca²⁺ are potential regulators of myosin-5 motor function, because they can destabilize its folded conformation. A combination of these regulators provides a versatile scheme in regulating myosin-5 motor function in the cell.     

Heavy and light roles: myosin in the morphogenesis of the heart

Myosin is an essential component of cardiac muscle, from the onset of cardiogenesis through to the adult heart. Although traditionally known for its role in energy transduction and force development, recent studies suggest that both myosin heavy-chain and myosin light-chain proteins are required for a correctly formed heart. Myosins are structural proteins that are not only expressed from early stages of heart development, but when mutated in humans they may give rise to congenital heart defects. This review will discuss the roles of myosin, specifically with regards to the developing heart. The expression of each myosin protein will be described, and the effects that altering expression has on the heart in embryogenesis in different animal models will be discussed. The human molecular genetics of the myosins will also be reviewed.     

Functions of actin in endocytosis

Endocytosis is a fundamental eukaryotic process required for remodelling plasma-membrane lipids and protein to ensure appropriate membrane composition. Increasing evidence from a number of cell types reveals that actin plays an active, and often essential, role at key endocytic stages. Much of our current mechanistic understanding of the endocytic process has come from studies in budding yeast and has been facilitated by yeast’s genetic amenability and by technological advances in live cell imaging. While endocytosis in metazoans is likely to be subject to a greater array of regulatory signals, recent reports indicate that spatiotemporal aspects of vesicle formation requiring actin are likely to be conserved across eukaryotic evolution. In this review we focus on the ‘modular’ model of endocytosis in yeast before highlighting comparisons with other cell types. Our discussion is limited to endocytosis involving clathrin as other types of endocytosis have not been demonstrated in yeast.     

Organization of myosin molecules in the muscle thick filament

Summary Tryptic treatment of muscle thick filaments reveals the underlying backbones of aggregatedl-meromyosin as a coli of 3 secondary filaments (helical repeat 130 nm) each in turn a coli of a 3 finer ones.     

Calorimetric studies on monomeric and polymeric actin

Differential scanning calorimetry of polymeric F-actin at pH 8.0 showed that the polymer had a concentration-independent thermal profile with a single transition temperature of 81 degrees C. In contrast, the thermal profile of G-actin was concentration-dependent, and although it resembled the F-actin profile at lower concentrations, it was found to have a more complex profile at higher protein concentrations.     

Calorimetric studies on monomeric and polymeric actin

Summary Differential scanning calorimetry of polymeric F-actin at pH 8.0 showed that the polymer had a concentration-independent thermal profile with a single transition temperature of 81 °C. In contrast, the thermal profile of G-actin was concentration-dependent, and although it resembled the F-actin profile at lower concentrations, it was found to have a more complex profile at higher protein concentrations.Acknowledgment. This research was carried out under grants to J.F.B. and P.J. from The Muscular Dystrophy Association of America.     

Espins and the actin cytoskeleton of hair cell stereocilia and sensory cell microvilli

The espins are novel actin-bundling proteins that are produced in multiple isoforms from a single gene. They are present at high concentration in the parallel actin bundle of hair cell stereocilia and are the target of deafness mutations in mice and humans. Espins are also enriched in the microvilli of taste receptor cells, solitary chemoreceptor cells, vomeronasal sensory neurons and Merkel cells, suggesting that espins play important roles in the microvillar projections of vertebrate sensory cells. Espins are potent actin-bundling proteins that are not inhibited by Ca2+. In cells, they efficiently elongate parallel actin bundles and, thereby, help determine the steadystate length of microvilli and stereocilia. Espins bind actin monomer via their WH2 domain and can assemble actin bundles in cells. Certain espin isoforms can also bind phosphatidylinositol 4,5-bisphosphate, profilins or SH3 proteins. These biological activities distinguish espins from other actin-bundling proteins and may make them well-suited to sensory cells.