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Spheroplasts (0):
Recent article
Spheroplasts
Microbiology (155)Pt 6 2009
Abstract
We have investigated the folding properties and cellular trafficking of a TolC derivative that lacks the entire signal sequence (TolCDelta2-22). A significant amount of TolCDelta2-22 was found to be soluble in the cytoplasm, and a fraction of it folded and trimerized into a conformation similar to t...
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PMID: 19383696
PDF is available here.
PDF is available here.
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Abstract
We report here on a new bacterial cell-based system, which is suitable for study of interactions between ligands and ligand-binding sites of eukaryotic Kv1.3 and Kv1.1 channels. To create this system, high-level expression of KcsA-Kv1.3 and KcsA-Kv1.1 hybrid proteins (ligand-binding sites of Kv1.3 o...
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PMID: 18649142
PDF is available here.
PDF is available here.
EMBO Journal (28)4 2009
Abstract
We show that during stress there is a dramatic change in the nucleosome organization of stress-responsive loci that depends on Hog1 and the RSC chromatin remodelling complex. Upon stress, the MAPK Hog1 physically interacts with RSC to direct its association with the ORF of osmo-responsive genes. In...
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PMID: 19153600
PDF is available here.
PDF is available here.
Abstract
We have developed a technology for the facile isolation of full-length IgG antibodies with desired specificity from combinatorial libraries expressed in Escherichia coli. Full-length heavy and light chains are expressed from a bicistronic operon and are secreted into the periplasm where they assembl...
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PMID: 19252859
PDF is available here.
PDF is available here.
FEMS Microbiology Letters (289)1 2008
Abstract
Peroxiredoxins are a class of peroxide-scavenging enzymes having a conserved cysteine residue(s) in their active centers. Thiol peroxidase (Tpx) is one of the peroxiredoxins identified in Escherichia coli. Despite the absence of the N-terminal signal sequence for transport across the membrane, it ha...
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PMID: 19054092
PDF is available here.
PDF is available here.
Biochemistry (New York) (73)11 2008
Abstract
Silver ions induce the efflux of potassium from cells of the yeast Saccharomyces cerevisiae but have no such effect on spheroplasts. Copper ions and the natural fungicide 2-O-3-hydroxyhexanoyl-beta-D-glucopyranosyl-(1-->4)-(6-O-acetyl-beta-D-glucopyranosyl-(1-->16)-2,15,16-trihydroxyhexadecanoic aci...
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PMID: 19120026
PDF is available here.
PDF is available here.
Journal of Biological Chemistry (283)40 2008
Abstract
We used yeast to investigate (i) the short term event(s) associated with the Crabtree effect and (ii) a putative role of hexose phosphates in the inhibition of respiration. Indeed, yeast divide into "Crabtree-positive," where the Crabtree effect occurs, and "Crabtree-negative," where it does not. In...
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PMID: 18682403
PDF is available here.
PDF is available here.
Journal of Biochemistry (143)6 2008
Abstract
A novel type of membrane vesicles was formed in vitro from microsomes of Saccharomyces cerevisiae, which carries Dpm1p, an enzyme involved in dolichol-sugar synthesis, but not a typical secretory cargo. While COPII vesicles formed in vitro were sedimentable by centrifugation at 200,000g(max) for 15...
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PMID: 18343875
PDF is available here.
PDF is available here.
Abstract
Genetic approaches have revealed more than 50 genes involved in the delivery of soluble zymogens like carboxypeptidase Y (CPY) to the lysosome-like vacuole in Saccharomyces cerevisiae. At least 20 of these genes function in transport between the prevacuolar endosome-like compartment (PVC) and the va...
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PMID: 19066018
PDF is available here.
PDF is available here.
Biochimica et Biophysica Acta (1778)1 2008
Abstract
We determined that HP(2-20) (residues 2-20 of parental HP derived from the N-terminus of Helicobacter pylori Ribosomal Protein L1) and its analogue, HPA3, exhibit broad-spectrum antimicrobial activity. The primary objective of the present study was to gain insight into the relevant mechanisms of act...
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PMID: 17961502
PDF is available here.
PDF is available here.
Journal of Lipid Research (48)11 2007
Abstract
We propose that Fat1p has a third region, which binds to the membrane and separates the highly conserved residues comprising the two halves of the ATP/AMP motif. The N(in)-C(in) topology results in the placement of the ATP/AMP and FATP/VLACS domains of Fat1p on the inner face of the plasma membrane....
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PMID: 17679730
PDF is available here.
PDF is available here.
Abstract
We have therefore explored the use of the OmpA signal sequence to deliberately target the expression of the murine CMP-sialic acid transporter, a Golgi-resident protein with 10 putative transmembrane domains, to the Escherichia coli inner membrane. Here, we show that the expression of an OmpA signal...
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PMID: 17764658
PDF is available here.
PDF is available here.
Ultramicroscopy (107)10-11 2007
Abstract
Atomic force microscopy (AFM) provides a unique opportunity to study live individual bacteria at the nanometer scale. In addition to providing accurate morphological information, AFM can be exploited to investigate membrane protein localization and molecular interactions on the surfa...
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PMID: 17574761
PDF is available here.
PDF is available here.
Journal of Virology (81)18 2007
Abstract
We have previously shown that the expression of human immunodeficiency virus type 1 (HIV-1) Gag protein in Saccharomyces cerevisiae spheroplasts produces Gag virus-like particles (VLPs) at the plasma membrane, indicating that yeast has all the host factors necessary for HIV-1 Gag assembly. Here we e...
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PMID: 17609278
PDF is available here.
PDF is available here.
Abstract
We show that scFv antibody fragments expressed in the APEx format allow the binding of spheroplasts to immobilized ligands. ScFv antibodies specific for the cardiac glycoside digoxin or for the protective antigen (PA) of Bacillus anthracis as a negative control were expressed in E. coli as fusions t...
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PMID: 17657772
PDF is available here.
PDF is available here.
Abstract
We studied internal initiation of translation in yeast using the dicistronic approach. Three viral sequences and a polylinker-derived reference sequence were inserted into the ICS of a dual reporter plasmid upstream of the firefly luciferase gene, luc. LUC expression was taken as a putative indicati...
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PMID: 17331675
PDF is available here.
PDF is available here.
Veterinary Microbiology (120)3-4 2007
Abstract
A Mycobacterium avium subspecies paratuberculosis (MAP) vaccine that reduced the incidence of clinical disease or reduced fecal shedding of MAP would aid control of Johne's disease (JD). The objective of the present study was to evaluate the efficacy of four MAP vaccine combinations, including cell-...
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PMID: 17123751
PDF is available here.
PDF is available here.
Abstract
Glucose uptake machinery was not affected in the presence of toxic metal ions in the case of high-resistant strains. But in the case of low Hg(2+)-resistant strain, glucose transport system may be affected either by inactivation of sensor proteins containing -SH group associated with glucose uptake....
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PMID: 17241342
PDF is available here.
PDF is available here.
Abstract
We present evidence that aquaporins can channel hydrogen peroxide (H2O2). Twenty-four aquaporins from plants and mammals were screened in five yeast strains differing in sensitivity toward oxidative stress. Expression of human AQP8 and plant Arabidopsis TIP1;1 and TIP1;2 in yeast decreased growth an...
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PMID: 17105724
PDF is available here.
PDF is available here.
Abstract
Several methods for DNA-mediated transformation of Pichia pastoris have been developed which vary in type of DNA that is transformable (e.g., linear versus circular) efficiency, cost, and labor and each is described in detail. As in Saccharomyces cerevisiae, gene replacement (also known as gene knoc...
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PMID: 17951633
PDF is available here.
PDF is available here.
Abstract
We describe a genomewide approach to classify all the cellular mRNAs according to their physical proximity with mitochondria; we also present real-time quantitative reverse transcription polymerase chain reaction monitoring of mRNA distribution to provide a quantified description of this localizatio...
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PMID: 18314748
PDF is available here.
PDF is available here.
Abstract
The complexes of repair DNA polymerase beta with 3'-exonuclease and some other proteins were isolated from the chromatin of hepatocytes of normal rats for the first time. Biopolymers were extracted from the chromatin by the solution of NaCl and Triton X-100. The extract was fractionated by gel-filtr...
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PMID: 18041131
PDF is available here.
PDF is available here.
Methods in Enzymology (422) 2007
Abstract
We employed a full-length EnvZ construct fused to the green fluorescent protein (GFP) that was overexpressed and targeted to the inner membrane. Spheroplasts were prepared and lysed in microtiter plates containing purified, fluorescent-labeled OmpR protein. Fluorescence resonance energy transfer (FR...
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PMID: 17628148
PDF is available here.
PDF is available here.
Abstract
This study compared the in-vitro properties and in-vivo effects of Escherichia coli filaments, spheroplasts and normal cells in a murine thigh infection model. E. coli was exposed to ceftazidime, meropenem or saline to obtain filaments, spheroplasts or normal bacilli, which were then injected into n...
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PMID: 17002610
PDF is available here.
PDF is available here.
Biophysical Journal (91)8 2006
Abstract
The mechanosensitive channel of small conductance (MscS) is a bacterial mechanosensitive channel that opens in response to rapid hypoosmotic stress. Since MscS can be opened solely by membrane stretch without help from any accessory protein, the lipid-protein interface must play a crucial role in se...
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PMID: 16861270
PDF is available here.
PDF is available here.
Abstract
We show that a novel prokaryotic voltage-gated K(+) (Kv) channel from Listeria monocytogenes (KvLm) embodies a rudimentary, yet robust, sensor sufficient to endow it with voltage-dependent features comparable to those of eukaryotic Kv channels. The most conspicuous feature of the KvLm sequence is th...
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PMID: 16908725
PDF is available here.
PDF is available here.
Abstract
We have previously reported that spheroplasts of Saccharomyces cerevisiae expressing HIV-1 Gag proteins from an episomal plasmid constitutively released a large amount of VLPs into culture media; however, commercially available ELISA kits which detect mature capsid of HIV-1 could not detect uncleave...
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PMID: 16807037
PDF is available here.
PDF is available here.
Abstract
We present methods to prepare infectious Sup35 protein aggregates and use them for genetic transformation of yeast. The protein aggregates are prepared from bacterially expressed recombinant protein, which is converted to amyloid fibers by extended incubation or by nucleated growth using yeast prion...
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PMID: 16759879
PDF is available here.
PDF is available here.
Abstract
We overexpressed this eukaryotic aquaporin in the methylotrophic yeast Pichia pastoris, and immunogold labeling of cellular cryosections showed that the protein accumulated in the plasma membrane, as well as vacuolar and other intracellular membranes. We then developed an in vivo functional assay fo...
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PMID: 16461705
PDF is available here.
PDF is available here.
Abstract
Species composition of micromycete complexes colonizing aspen, birch, and pine wood was studied. Calculation of the Sorens-Tchekanovsky similarity coefficients showed that these complexes shared a high degree of similarity. They were dominated by the representatives of the genera Penicillium, Paecil...
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PMID: 16521580
PDF is available here.
PDF is available here.
Abstract
The effect of microdoses of copper ions bound with the external surface of the cytoplasmic membrane of Escherichia coli spheroplasts on the charge density of its superficial structures was investigated by the ESR method. Positively charged spin probes of KAT(n)-type and the newly synthesized KAT15 w...
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PMID: 16521558
PDF is available here.
PDF is available here.
Microbiology (151)Pt 12 2005
Abstract
Acetobacter tropicalis SKU1100 produces a pellicle polysaccharide, consisting of galactose, glucose and rhamnose, which attaches to the cell surface. This strain forms two types of colony on agar plates: a rough-surfaced colony (R strain) and a mucoid smooth-surfaced colony (S strain). The R strain...
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PMID: 16339956
PDF is available here.
PDF is available here.
Helicobacter (10)6 2005
Abstract
This study demonstrated the presence of the peptidergic autolytic substances in the culture supernatant of H. pylori KZ109 cells. The results of this study should be useful for further studies aimed at elucidation of the strategy of survival of H. pylori in the gastric environment and elucidation of...
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PMID: 16302982
PDF is available here.
PDF is available here.
Ultramicroscopy (105)1-4 2005
Abstract
The cytoplasmic membrane of Escherichia coli (E. coli) is the location of numerous, chemically specific transporters and recognition elements. Investigation of this membrane in vivo by atomic force microscopy (AFM) requires removal of the cell wall and stable immobilization of the spheroplast. AFM i...
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PMID: 16112809
PDF is available here.
PDF is available here.
Biochemistry (New York) (70)9 2005
Abstract
The effect of cultivation time and concentration of inorganic phosphate (P(i)) in the culture medium on the accumulation of polyphosphates (polyP) and the activity of two cytosolic exopolyphosphatases of the yeast Saccharomyces cerevisiae was studied: an exopolyphosphatase of 40 kD encoded by PPX1 a...
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PMID: 16266267
PDF is available here.
PDF is available here.
Abstract
Permeable spheroplasts were prepared from two strains of Saccharomyces cerevisiae by incubating with zymolyase without a permeabilizing agent. The loss of the plasma membrane barrier was confirmed by the nucleotide release, the activity of glucose 6-phosphate dehydrogenase with external substrates a...
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PMID: 16034802
PDF is available here.
PDF is available here.
Abstract
We examined the effects of ethyl and propyl paraben, on gating of the E. coli mechanosensitive channel of large conductance (MscL) reconstituted into azolectin liposomes. We found that propyl and ethyl paraben spontaneously activate MscL. Moreover, the addition of propyl paraben caused an increase i...
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PMID: 15770478
PDF is available here.
PDF is available here.
Abstract
We present a method to visualise MscL in living bacteria using confocal microscopy. By creating a gene fusion between mscl and the gene encoding the green fluorescent protein (GFP) we were able to express the fusion protein MscL-GFP in bacteria. We show that MscL-GFP is present in the cytoplasmic me...
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PMID: 15812637
PDF is available here.
PDF is available here.
Abstract
We have investigated the effect of high hydrostatic pressure on MscS, the bacterial mechanosensitive channel of small conductance. Pressure affected channel kinetics but not conductance. At negative pipette voltages (corresponding to membrane depolarization in the inside-out patch configuration used...
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PMID: 15834558
PDF is available here.
PDF is available here.
Abstract
We investigated the possibility that LPS transport proceeds via a soluble intermediate associated with a periplasmic chaperone analogous to the Lol-dependent transport mechanism of lipoproteins. Whereas newly synthesized lipoproteins could be released from spheroplasts of Escherichia coli upon addit...
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PMID: 15576375
PDF is available here.
PDF is available here.
Abstract
We characterized the gating of MscS in Escherichia coli spheroplasts in a triple mutant (mscL-, mscS-, mscK-) background. We used a pressure clamp apparatus along with software developed in-lab to generate dose-response curves directly from two-channel recordings of current and pressure. In contrast...
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PMID: 15657299
PDF is available here.
PDF is available here.
Abstract
We present thermodynamic analysis of several GOF mutants to clarify the nature and position of low-conducting conformations in the transition pathway. Unlike wild-type (WT) MscL, which predominantly occupies the closed and fully open states with very brief substates, the mild V23T GOF mutant frequen...
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PMID: 15684093
PDF is available here.
PDF is available here.
Abstract
Zygosaccharomyces bailii is a commercially important spoilage yeast capable of growth at low pH in the presence of weak organic acid preservatives, such as benzoic acid. A patch-clamp electrophysiological analysis of plasma membrane K+ transport revealed a high conductance pathway for low-affinity K...
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PMID: 15580608
PDF is available here.
PDF is available here.
Abstract
We developed rapid-freeze and deep-etch techniques to image the yeast cell cortex in spheroplasted cells at high resolution. The cortical actin cytoskeleton assembles into conical or mound-like structures composed of short, cross-linked filaments. The Arp2/3 complex localizes near the apex of these...
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PMID: 15525671
PDF is available here.
PDF is available here.
Biochemistry (Washington) (43)47 2004
Abstract
The phospholipid composition of Rhodobacter sphaeroides cells resuspended in various hypertonic solutions has been examined by thin-layer chromatography and ESI mass spectrometry. R. sphaeroides responds to hyperosmotic stress by increasing the amount of cardiolipin in the membranes; this phenomenon...
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PMID: 15554714
PDF is available here.
PDF is available here.
Abstract
We have previously reported that a novel S-benzylisothiourea derivative, S-(3,4-dichlorobenzyl)isothiourea, tentatively named A22, induced spherical cells in Escherichia coli. To elucidate the structural element(s) required for inducing these spherical cells, the biological activity of S-benzylisoth...
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PMID: 15564663
PDF is available here.
PDF is available here.
FEMS Yeast Research (5)2 2004
Abstract
We show that additional deletion of YCA1, coding for the yeast metacaspase, suppressed the ubp10 disruptant phenotype. Moreover, YCA1 overexpression, without any external stimulus, had a detrimental effect on growth and viability of ubp10 cells accompanied by an increase of apoptotic cells. This res...
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PMID: 15489197
PDF is available here.
PDF is available here.
Biophysical Journal (87)5 2004
Abstract
We probe the relative position of specific residues predicted to line the pore of MscL in either the closed state or during the closed-to-open transition by engineering single-site histidine substitutions and assessing the ability of Ni2+, Cd2+ or Zn2+ ions to affect channel activity. All residues p...
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PMID: 15339809
PDF is available here.
PDF is available here.
Abstract
The respiration, membrane potential (Deltapsi), and oxidative phosphorylation of mitochondria in situ were determined in spheroplasts obtained from Candida albicans control strain ATCC 90028 by lyticase treatment. Mitochondria in situ were able to phosphorylate externally added ADP (200 microM) in t...
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PMID: 15448865
PDF is available here.
PDF is available here.
FEMS Yeast Research (4)8 2004
Abstract
Study of Saccharomyces cerevisiae killer toxin-sensitive strains with the deltakre2 phenotype (resistant to toxin K1, sensitive to toxin K2) showed that the phenotype is complemented by the KRE2 gene not only in intact cells but also in spheroplasts, and resistance to K1 thus resides very probably i...
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PMID: 15450187
PDF is available here.
PDF is available here.
Abstract
We examined the importance of membrane ergosterol and sphingolipids in the drug susceptibilities of Candida albicans. We used three independent methods to test the drug susceptibilities of erg mutant cells, which were defective in ergosterol biosynthesis. While spot and filter disk assays revealed t...
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PMID: 15105135
PDF is available here.
PDF is available here.
Abstract
These results suggest that low-strength conditions and the cellular metabolic state may modulate the candidacidal activity of human lactoferrin....
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PMID: 15047526
PDF is available here.
PDF is available here.
Journal of Biological Chemistry (279)13 2004
Abstract
We report on the synthesis, biological function, and a plausible mode of action of a new group of lipopeptides with potent antifungal and antibacterial activities. These lipopeptides are derived from positively charged peptides containing d- and l-amino acids (diastereomers) that are palmitoylated (...
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PMID: 14709550
PDF is available here.
PDF is available here.
FEMS Yeast Research (4)6 2004
Abstract
Isolated mitochondria of Saccharomyces cerevisiae cells grown on glucose possess acid-soluble inorganic polyphosphate (polyP). Its level strongly depends on phosphate (P(i)) concentration in the culture medium. The polyP level in mitochondria showed 11-fold decrease under 0.8 mM P(i) as compared wit...
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PMID: 15040953
PDF is available here.
PDF is available here.
Decrease of H2O2 plasma membrane permeability during adaptation to H2O2 in Saccharomyces cerevisiae.
Abstract
We extended these studies and tested whether an active regulation of biomembranes permeability characteristics is part of the cell response to oxidative stress. Using Saccharomyces cerevisiae as a model, we showed that: (a) H2O2 gradients across the plasma membrane are formed upon exposure to extern...
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PMID: 14645222
PDF is available here.
PDF is available here.
Abstract
We describe a novel activity of DsbB to induce an electronic transition of the bound ubiquinone molecule. This transition was characterized by a striking emergence of an absorbance peak at 500 nm giving rise to a visible pink color. The ubiquinone red-shift was observed stably for the DsbA(C33S)-Dsb...
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PMID: 14634016
PDF is available here.
PDF is available here.
Microbiological Research (159)1 2004
Abstract
To emphasise the role of outer and inner membranes in the resistance of Pseudomonas aeruginosa to bactericidal activity of various disinfectants, spheroplasts and whole cells were compared. Spheroplasts are more sensitive than whole cells to quaternary ammonium compounds such as didecyl dimethyl amm...
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PMID: 15160607
PDF is available here.
PDF is available here.
Abstract
We aimed to identify the substrate for this transporter. Cells that constitutively express the yiaK-S gene cluster metabolized the rare pentose L-xylulose, while deletion of the yiaMNO transporter genes reduced L-xylulose metabolism. The periplasmic substrate-binding protein YiaO was found to bind L...
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PMID: 14668138
PDF is available here.
PDF is available here.
Abstract
Antonie Van Leeuwenhoek (85)1 2004
Abstract
Under anaerobiosis, the mitochondrion of Saccharomyces cerevisiae is restricted to unstructured promitochondria. These promitochondria provide unknown metabolic functions that are required for growth. Since high glucose concentrations are mainly fermented by S. cerevisiae during stationary phase (du...
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PMID: 15031659
PDF is available here.
PDF is available here.