The Plasma Membrane of Saccharomyces cerevisiae:Structure, Function, and Biogenesis

The composition of phospholipids, sphingolipids, and sterols in the plasma membrane has a strong influence on the activity of the proteins associated or embedded in the lipid bilayer. Since most lipid-synthesizing enzymes in Saccharomyces cerevisiae are located in intracellular organelles, an extensive pur of lipids fi om these organelles to the plasma membrane is required. Although the pathway of protein traffic to the plasma membrane is similar to that of most of the lipids, the bulk flow of lipids is separate from vesicle-mediated protein transport. Recent advances in the analysis of membrane budding and membrane fusion indicate that the mechanisms of protein transport from the endoplasmic reticulum to the Golgi and fi om the Golgi to plasma membrane are similar. The majority of plasma membrane proteins transport solutes across the membrane. A number of ATP-dependent export systems have been detected that couple the hydrolysis of ATP to transport of molecules our of the cell. The hydrolysis of ATP by the plasma membrane H+-ATPase generates a proton motive force which is used to drive secondary transport-processes. In S. cerevisiae, mal Ey substrates are transported by more than one system. Transport of monosaccharide is catalyzed by uniport systems, while transport of disaccharides, amino acids, and nucleosides is mediated by proton symport systems. Transport activity can be regulated at the level of transcription, e.g., induction and (catabolite) repression but transport proteins can also be affected posttranslationally by a process termed catabolite inactivation. Catabolite inactivation is triggered by the addition of fermentable sugars, intracellular acidification, stress conditions, and/or nitrogen starvation. Phosphorylation and/or ubiquitination of the transport proteins has been proposed as an initial step in the controlled inactivation and regulation of the target enzyme. The use of artificial membranes, like secretory vesicles and plasma membranes fused with proteoliposomes, as model systems for studies on the mechanism and regulation of transport is evaluated.</p

Measurement of Low-Energy Cosmic-Ray Antiprotons at Solar Minimum

The absolute fluxes of the cosmic-ray antiprotons at solar minimum are measured in the energy range 0.18 to 1.4 GeV, based on 43 events unambiguously detected in BESS '95 data. The resultant energy spectrum appears to be flat below 1 GeV, compatible with a possible admixture of primary antiproton component with a soft energy spectrum, while the possibility of secondary antiprotons alone explaining the data cannot be excluded with the present accuracy. Further improvement of statistical accuracy and extension of the energy range are planned in future BESS flights.Comment: REVTeX, 4 pages including 4 eps figures. Submitted to PR

Structure and function of the bacterial heterodimeric ABC transporter CydDC: stimulation of ATPase activity by thiol and heme compounds.

In Escherichia coli, the biogenesis of both cytochrome bd-type quinol oxidases and periplasmic cytochromes requires the ATP-binding cassette-type cysteine/GSH transporter, CydDC. Recombinant CydDC was purified as a heterodimer and found to be an active ATPase both in soluble form with detergent and when reconstituted into a lipid environment. Two-dimensional crystals of CydDC were analyzed by electron cryomicroscopy, and the protein was shown to be made up of two non-identical domains corresponding to the putative CydD and CydC subunits, with dimensions characteristic of other ATP-binding cassette transporters. CydDC binds heme b. Detergent-solubilized CydDC appears to adopt at least two structural states, each associated with a characteristic level of bound heme. The purified protein in detergent showed a weak basal ATPase activity (approximately 100 nmol Pi/min/mg) that was stimulated ∼3-fold by various thiol compounds, suggesting that CydDC could act as a thiol transporter. The presence of heme (either intrinsic or added in the form of hemin) led to a further enhancement of thiol-stimulated ATPase activity, although a large excess of heme inhibited activity. Similar responses of the ATPase activity were observed with CydDC reconstituted into E. coli lipids. These results suggest that heme may have a regulatory role in CydDC-mediated transmembrane thiol transport

Cys34-cysteinylated human serum albumin is a sensitive plasma marker in oxidative stress-related chronic diseases

The degree of oxidized cysteine (Cys) 34 in human serum albumin (HSA), as determined by high performance liquid chromatography (HPLC), is correlated with oxidative stress related pathological conditions. In order to further characterize the oxidation of Cys34-HSA at the molecular level and to develop a suitable analytical method for a rapid and sensitive clinical laboratory analysis, the use of electrospray ionization time-of-flight mass spectrometer (ESI-TOFMS) was evaluated. A marked increase in the cysteinylation of Cys34 occurs in chronic liver and kidney diseases and diabetes mellitus. A significant positive correlation was observed between the Cys-Cys34-HSA fraction of plasma samples obtained from 229 patients, as determined by ESI-TOFMS, and the degree of oxidized Cys34-HSA determined by HPLC. The Cys-Cys34-HSA fraction was significantly increased with the progression of liver cirrhosis, and was reduced by branched chain amino acids (BCAA) treatment. The changes in the Cys-Cys34-HSA fraction were significantly correlated with the alternations of the plasma levels of advanced oxidized protein products, an oxidative stress marker for proteins. The binding ability of endogenous substances (bilirubin and tryptophan) and drugs (warfarin and diazepam) to HSA purified from chronic liver disease patients were significantly suppressed but significantly improved by BCAA supplementation. Interestingly, the changes in this physiological function of HSA in chronic liver disease were correlated with the Cys-Cys34-HSA fraction. In conclusion, ESI-TOFMS is a suitable high throughput method for the rapid and sensitive quantification of Cys-Cys34-HSA in a large number of samples for evaluating oxidative stress related chronic disease progression or in response to a treatment

Precision Measurement of Cosmic-Ray Antiproton Spectrum

The energy spectrum of cosmic-ray antiprotons has been measured in the range 0.18 to 3.56 GeV, based on 458 antiprotons collected by BESS in recent solar-minimum period. We have detected for the first time a distinctive peak at 2 GeV of antiprotons originating from cosmic-ray interactions with the interstellar gas. The peak spectrum is reproduced by theoretical calculations, implying that the propagation models are basically correct and that different cosmic-ray species undergo a universal propagation. Future BESS flights toward the solar maximum will help us to study the solar modulation and the propagation in detail and to search for primary antiproton components.Comment: REVTeX, 4 pages including 4 eps figure

Measurements of Proton, Helium and Muon Spectra at Small Atmospheric Depths with the BESS Spectrometer

The cosmic-ray proton, helium, and muon spectra at small atmospheric depths of 4.5 -- 28 g/cm^2 were precisely measured during the slow descending period of the BESS-2001 balloon flight. The variation of atmospheric secondary particle fluxes as a function of atmospheric depth provides fundamental information to study hadronic interactions of the primary cosmic rays with the atmosphere.Comment: 21 pages, 11 figures, 4 table

Measurements of Primary and Atmospheric Cosmic-Ray Spectra with the BESS-TeV Spectrometer

Primary and atmospheric cosmic-ray spectra were precisely measured with the BESS-TeV spectrometer. The spectrometer was upgraded from BESS-98 to achieve seven times higher resolution in momentum measurement. We report absolute fluxes of primary protons and helium nuclei in the energy ranges, 1-540 GeV and 1-250 GeV/n, respectively, and absolute flux of atmospheric muons in the momentum range 0.6-400 GeV/c.Comment: 26 pages, 9 figures, 3 tables, Submitted to Phys. Lett.

Precise Measurements of Atmospheric Muon Fluxes with the BESS Spectrometer

The vertical absolute fluxes of atmospheric muons and muon charge ratio have been measured precisely at different geomagnetic locations by using the BESS spectrometer. The observations had been performed at sea level (30 m above sea level) in Tsukuba, Japan, and at 360 m above sea level in Lynn Lake, Canada. The vertical cutoff rigidities in Tsukuba (36.2 N, 140.1 E) and in Lynn Lake (56.5 N, 101.0 W) are 11.4 GV and 0.4 GV, respectively. We have obtained vertical fluxes of positive and negative muons in a momentum range from 0.6 to 20 GeV/c with systematic errors less than 3 % in both measurements. By comparing the data collected at two different geomagnetic latitudes, we have seen an effect of cutoff rigidity. The dependence on the atmospheric pressure and temperature, and the solar modulation effect have been also clearly observed. We also clearly observed the decrease of charge ratio of muons at low momentum side with at higher cutoff rigidity region.Comment: 35 pages, 9 figures. Submitted to Astroparticle Physic