36 research outputs found

    Binding of 25-hydroxycholesterol and cholesterol to different cytoplasmic proteins

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    Linear Collider Capabilities for Supersymmetry in Dark Matter Allowed Regions of the mSUGRA Model

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    Recent comparisons of minimal supergravity (mSUGRA) model predictions with WMAP measurements of the neutralino relic density point to preferred regions of model parameter space. We investigate the reach of linear colliders (LC) with s=0.5\sqrt{s}=0.5 and 1 TeV for SUSY in the framework of the mSUGRA model. We find that LCs can cover the entire stau co-annihilation region provided \tan\beta \alt 30. In the hyperbolic branch/focus point (HB/FP) region of parameter space, specialized cuts are suggested to increase the reach in this important ``dark matter allowed'' area. In the case of the HB/FP region, the reach of a LC extends well past the reach of the CERN LHC. We examine a case study in the HB/FP region, and show that the MSSM parameters μ\mu and M2M_2 can be sufficiently well-measured to demonstrate that one would indeed be in the HB/FP region, where the lightest chargino and neutralino have a substantial higgsino component.Comment: 29 pages, 15 EPS figures; updated version slightly modified to conform with published versio

    The Gravitational Universe

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    The last century has seen enormous progress in our understanding of the Universe. We know the life cycles of stars, the structure of galaxies, the remnants of the big bang, and have a general understanding of how the Universe evolved. We have come remarkably far using electromagnetic radiation as our tool for observing the Universe. However, gravity is the engine behind many of the processes in the Universe, and much of its action is dark. Opening a gravitational window on the Universe will let us go further than any alternative. Gravity has its own messenger: Gravitational waves, ripples in the fabric of spacetime. They travel essentially undisturbed and let us peer deep into the formation of the first seed black holes, exploring redshifts as large as z ~ 20, prior to the epoch of cosmic re-ionisation. Exquisite and unprecedented measurements of black hole masses and spins will make it possible to trace the history of black holes across all stages of galaxy evolution, and at the same time constrain any deviation from the Kerr metric of General Relativity. eLISA will be the first ever mission to study the entire Universe with gravitational waves. eLISA is an all-sky monitor and will offer a wide view of a dynamic cosmos using gravitational waves as new and unique messengers to unveil The Gravitational Universe. It provides the closest ever view of the early processes at TeV energies, has guaranteed sources in the form of verification binaries in the Milky Way, and can probe the entire Universe, from its smallest scales around singularities and black holes, all the way to cosmological dimensions

    Assay of oxysterol-binding protein in a mouse fibroblast, cell-free system. Dissociation constant and other properties of the system.

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    Procedures for determining a 7.5 S oxysterol-binding protein in the cytosol fraction of cultured mouse fibroblasts were developed. The methods involved precipitation of cytosolic proteins between 0.3 and 0.4 saturation with (NH4)2SO4, incubation of the proteins with 25-hydroxy[3H]cholesterol at 0 degrees C and analysis by velocity sedimentation of 7.5 S radioactivity or of specific binding using dextran-charcoal to adsorb free sterol. By these means it was shown that binding of the ligand to the protein in a citric acid-phosphate buffer was optimal at pH 5.5 and that the sedimentation rate of the complex was greater at pH 7.4 (7.5 S) than at pH 5.5 (6.9 S). The binding protein was essentially saturated at a diol concentration of about 20 X 10(-9) M. The apparent Kd of the sterol-protein complex was approximately 3.9 X 10(-9) M. Cholesterol did not bind to 25-hydroxycholesterol-binding sites on the 7.5 S protein, whereas several oxysterols that are potent suppressors of 3-hydroxy-3-methylglutaryl CoA reductase also inhibited the binding of 25-hydroxycholesterol. One of these sterols, 5 alpha-cholest-8(14)-en-3 beta-ol-15-one was shown to compete for sites occupied by 25-hydroxycholesterol

    Sex reversal in C57BL/6J-YPOS mice corrected by a Sry transgene.

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    C57BL/6J mice carrying a Mus domesticus poschiavinus Y chromosome (YPOS) develop as females with ovarian tissue or as hermaphrodites with ovarian and testicular tissue. We tested the hypothesis that the Y-linked component of this inherited sex reversal is caused by the M. d. poschiavinus Y-linked testis determining gene (symbolized Tdy or Sry) by examining gonadal development in C57BL/6J XYPOS mice carrying a M. musculus allele of Sry as a transgene. We found that in the presence of the transgene, XYPOS mice developed exclusively testicular tissue. This result indicates that the Sry allele carried on the YPOS chromosome is responsible for development of ovarian tissue in the C57BL/6J inbred strain background. We discuss this finding in light of DNA polymorphisms present in Sry alleles carried by various M. domesticus and M. musculus Y chromosomes. In addition, we present a hypothesis concerning the timing of expression of the testicular and ovarian determining genes in the developing fetal gonad based on the organization of ovarian and testicular tissue in ovotestes

    A proteolytic fragment of the oxysterol receptor which retains oxysterol binding activity.

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    The structural organization of the oxysterol receptor, postulated to be involved in the regulation of 3-hydroxy-3-methylglutaryl CoA reductase and cholesterol biosynthesis in mammalian cells, has been explored by limited proteolysis with trypsin, alpha-chymotrypsin, and endoproteinase GluC. Treatment with each of these proteases converts the receptor from a homodimer of approximately 95 kDa subunits to a 44-kDa form, based on hydrodynamic measurements by sucrose density gradient centrifugation and gel filtration chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of photoaffinity-labeled preparations indicates that the oxysterol binding site is on a 28-kDa fragment within the 44-kDa limit form of the receptor. The limit proteolytc form exhibits the high affinity and structural specificity for oxysterols of the native dimeric receptor with an increase in the rate constant of association for 25-hydroxycholesterol. The proteolytic form also shows an increased binding affinity for nonspecific DNA, but no sequence specificity for the oxysterol regulatory element from the reductase gene was detected

    Different forms of the oxysterol-binding protein. Binding kinetics and stability.

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    Based upon measurements of the sedimentation coefficient and the Stokes radii, three forms of the oxysterol-binding protein were identified. The unliganded binding protein was the largest (7.7 S, Stokes radius = 71.6 A, Mr = 236,000) was relatively asymmetric (f/f0 = 1.7), and was composed of at least three subunits. Binding of 25-hydroxycholesterol was associated with a reduction in the size of the protein (7.5 S, Stokes radius = 50 A, Mr approximately 169,000) and an increase in symmetry (f/f0 = 1.4), due to the loss of a subunit of Mr approximately 67,000. At pH 6 or lower, the Mr = 169,000 sterol-protein complex was altered so that reversible dissociation to give a smaller (4.2 S, Stokes radius = 53 A, Mr = 97,000) more asymmetric (f/f0 = 1.8) sterol-protein complex occurred when it was sedimented in a sucrose gradient buffered at pH 7.4 containing 0.3 M KCl and 2.5 M urea. Irreversible dissociation of the 7.5 S, Mr = 169,000 form to a 4.2 S form occurred spontaneously when the complex in whole cytosol buffered at pH 7.8 was allowed to stand overnight at 0 degree C, or when the partially purified complex was incubated at pH 5.5 at 0 degree C for several days. The partially purified, unliganded binding protein was unstable at 0 degree C (approximately 75% loss of binding activity in 24 h) whereas the liganded protein was stable for 7 days at 0 degree C although irreversible conversion to a 4.2 S form occurred under some conditions. Rates of sterol binding and dissociation were increased in the presence of 2.5 M urea at pH 7.4 or when the pH was lowered to 5.5 Kd values were not greatly altered under the various incubation conditions
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