Bead movement by single kinesin molecules studied with optical tweezers

Kinesin, a mechanoenzyme that couples ATP hydrolysis to movement along microtubules, is thought to power vesicle transport and other forms of microtubule-based motility. Here, microscopic silica beads were precoated with carrier protein, exposed to low concentrations of kinesin, and individually manipulated with a single-beam gradient-force optical particle trap ('optical tweezers') directly onto microtubules. Optical tweezers greatly improved the efficiency of the bead assay, particularly at the lowest kinesin concentrations (corresponding to approximately 1 molecule per bead). Beads incubated with excess kinesin moved smoothly along a microtubule for many micrometres, but beads carrying from 0.17-3 kinesin molecules per bead, moved, on average, only about 1.4 microns and then spontaneously released from the microtubule. Application of the optical trap directly behind such moving beads often pulled them off the microtubule and back into the centre of the trap. This did not occur when a bead was bound by an AMP.PNP-induced rigor linkage, or when beads were propelled by several kinesin molecules. Our results are consistent with a model in which kinesin detaches briefly from the microtubule during a part of each mechanochemical cycle, rather than a model in which kinesin remains bound at all times.     

Origin of a 'Southern Hemisphere' geochemical signature in the Arctic upper mantle

The Gakkel ridge, which extends under the Arctic ice cap for approximately 1,800 km, is the slowest spreading ocean ridge on Earth. Its spreading created the Eurasian basin, which is isolated from the rest of the oceanic mantle by North America, Eurasia and the Lomonosov ridge. The Gakkel ridge thus provides unique opportunities to investigate the composition of the sub-Arctic mantle and mantle heterogeneity and melting at the lower limits of seafloor spreading. The first results of the 2001 Arctic Mid-Ocean Ridge Expedition (ref. 1) divided the Gakkel ridge into three tectonic segments, composed of robust western and eastern volcanic zones separated by a 'sparsely magmatic zone'. On the basis of Sr-Nd-Pb isotope ratios and trace elements in basalts from the spreading axis, we show that the sparsely magmatic zone contains an abrupt mantle compositional boundary. Basalts to the west of the boundary display affinities to the Southern Hemisphere 'Dupal' isotopic province, whereas those to the east-closest to the Eurasian continent and where the spreading rate is slowest-display affinities to 'Northern Hemisphere' ridges. The western zone is the only known spreading ridge outside the Southern Hemisphere that samples a significant upper-mantle region with Dupal-like characteristics. Although the cause of Dupal mantle has been long debated, we show that the source of this signature beneath the western Gakkel ridge was subcontinental lithospheric mantle that delaminated and became integrated into the convecting Arctic asthenosphere. This occurred as North Atlantic mantle propagated north into the Arctic during the separation of Svalbard and Greenland.     

Action of group A streptococcus extracellular product(s) on the connective tissue of the human heart valve

Summary The cultivation of a rheumatogenic strain of group A streptococcus in presence of human heart valve connective tissues leads to the release of substance(s) reacting with antistreptococcal polysaccharide and antihuman glycoprotein antibodies.     

Tethys and Dione as sources of outward-flowing plasma in Saturn's magnetosphere

Rotating at over twice the angular speed of Earth, Saturn imposes a rapid spin on its magnetosphere. As a result, cold, dense plasma is believed to be flung outward from the inner magnetosphere by centrifugal force and replaced by hotter, more tenuous plasma from the outer magnetosphere. The centrifugal interchange of plasmas in rotating magnetospheres was predicted many years ago and was conclusively demonstrated by observations in Jupiter's magnetosphere, which--like that of Saturn (but unlike that of Earth)--is rotationally dominated. Recent observations in Saturn's magnetosphere have revealed narrow injections of hot, tenuous plasma believed to be the inward-moving portion of the centrifugal interchange cycle. Here we report observations of the distribution of the angle between the electron velocity vector and the magnetic field vector ('pitch angle') obtained in the cold, dense plasma adjacent to these inward injection regions. The observed pitch-angle distributions are indicative of outward plasma flow and consistent with centrifugal interchange in Saturn's magnetosphere. Further, we conclude that the observed double-peaked ('butterfly') pitch-angle distributions result from the transport of plasma from regions near the orbits of Dione and Tethys, supporting the idea of distinct plasma tori associated with these moons.     

Coated pits, coated vesicles, and receptor-mediated endocytosis.

Proteins and peptides can enter cells by receptor-mediated endocytosis, a coupled process by which selected extracellular proteins or peptides are first bound to specific cell surface receptors and then rapidly internalised by the cell. Internalisation follows clustering of the receptors in specialised regions of the cell surface called coated pits that invaginate to form intracellular coated vesicles. It is now recognised that receptor-mediated endocytosis has a fundamental role in the growth, nutrition and differentiation of animal cells.     

Structure of the insulin receptor substrate IRS-1 defines a unique signal transduction protein.

Since the discovery of insulin nearly 70 years ago, there has been no problem more fundamental to diabetes research than understanding how insulin works at the cellular level. Insulin binds to the alpha subunit of the insulin receptor which activates the tyrosine kinase in the beta subunit, but the molecular events linking the receptor kinase to insulin-sensitive enzymes and transport processes are unknown. Our discovery that insulin stimulates tyrosine phosphorylation of a protein of relative molecular mass between 165,000 and 185,000, collectively called pp185, showed that the insulin receptor kinase has specific cellular substrates. The pp185 is a minor cytoplasmic phosphoprotein found in most cells and tissues; its phosphorylation is decreased in cells expressing mutant receptors defective in signalling. We have now cloned IRS-1, which encodes a component of the pp185 band. IRS-1 contains over ten potential tyrosine phosphorylation sites, six of which are in Tyr-Met-X-Met motifs. During insulin stimulation, the IRS-1 protein undergoes tyrosine phosphorylation and binds phosphatidylinositol 3-kinase, suggesting that IRS-1 acts as a multisite 'docking' protein to bind signal-transducing molecules containing Src-homology 2 and Src-homology-3 domains. Thus IRS-1 may link the insulin receptor kinase and enzymes regulating cellular growth and metabolism.