19 research outputs found
Transcriptional and Proteomic Analysis of a Ferric Uptake Regulator (Fur) Mutant of Shewanella oneidensis: Possible Involvement of Fur in Energy Metabolism, Transcriptional Regulation, and Oxidative Stress
The iron-directed, coordinate regulation of genes depends on the fur (ferric uptake regulator) gene product, which acts as an iron-responsive, transcriptional repressor protein. To investigate the biological function of a fur homolog in the dissimilatory metal-reducing bacterium Shewanella oneidensis MR-1, a fur knockout strain (FUR1) was generated by suicide plasmid integration into this gene and characterized using phenotype assays, DNA microarrays containing 691 arrayed genes, and two-dimensional polyacrylamide gel electrophoresis. Physiological studies indicated that FUR1 was similar to the wild-type strain when they were compared for anaerobic growth and reduction of various electron acceptors. Transcription profiling, however, revealed that genes with predicted functions in electron transport, energy metabolism, transcriptional regulation, and oxidative stress protection were either repressed (ccoNQ, etrA, cytochrome b and c maturation-encoding genes, qor, yiaY, sodB, rpoH, phoB, and chvI) or induced (yggW, pdhC, prpC, aceE, fdhD, and ppc) in the fur mutant. Disruption of fur also resulted in derepression of genes (hxuC, alcC, fhuA, hemR, irgA, and ompW) putatively involved in iron uptake. This agreed with the finding that the fur mutant produced threefold-higher levels of siderophore than the wild-type strain under conditions of sufficient iron. Analysis of a subset of the FUR1 proteome (i.e., primarily soluble cytoplasmic and periplasmic proteins) indicated that 11 major protein species reproducibly showed significant (P < 0.05) differences in abundance relative to the wild type. Protein identification using mass spectrometry indicated that the expression of two of these proteins (SodB and AlcC) correlated with the microarray data. These results suggest a possible regulatory role of S. oneidensis MR-1 Fur in energy metabolism that extends the traditional model of Fur as a negative regulator of iron acquisition systems
Identification of Membrane Proteins in the Hyperthermophilic Archaeon Pyrococcus Furiosus Using Proteomics and Prediction Programs
Cell-free extracts from the hyperthermophilic archaeon Pyrococcus furiosus were
separated into membrane and cytoplasmic fractions and each was analyzed by 2D-gel
electrophoresis. A total of 66 proteins were identified, 32 in the membrane fraction and 34
in the cytoplasmic fraction. Six prediction programs were used to predict the subcellular
locations of these proteins. Three were based on signal-peptides (SignalP, TargetP, and
SOSUISignal) and three on transmembrane-spanning α-helices (TSEG, SOSUI, and
PRED-TMR2). A consensus of the six programs predicted that 23 of the 32 proteins
(72%) from the membrane fraction should be in the membrane and that all of the proteins
from the cytoplasmic fraction should be in the cytoplasm. Two membrane-associated
proteins predicted to be cytoplasmic by the programs are also predicted to consist
primarily of transmembrane-spanning β-sheets using porin protein models, suggesting that
they are, in fact, membrane components. An ATPase subunit homolog found in the
membrane fraction, although predicted to be cytoplasmic, is most likely complexed with
other ATPase subunits in the membrane fraction. An additional three proteins predicted to
be cytoplasmic but found in the membrane fraction, may be cytoplasmic contaminants.
These include a chaperone homolog that may have attached to denatured membrane
proteins during cell fractionation. Omitting these three proteins would boost the
membrane-protein predictability of the models to near 80%. A consensus prediction using
all six programs for all 2242 ORFs in the P. furiosus genome estimates that 24% of the
ORF products are found in the membrane. However, this is likely to be a minimum value
due to the programs’ inability to recognize certain membrane-related proteins, such as
subunits associated with membrane complexes and porin-type proteins
Energetics studies of organic and biomolecular ions by surface-induced dissociation.
Energetics studies of organic and biomolecular ions by surface-induced dissociation
Relative Dissociation Energies of Protonated Peptides by Electrospray Ionization/Surface-Induced Dissociation
Soft Coulomb gap and asymmetric scaling towards metal-insulator quantum criticality in multilayer MoS2
Quantum localization-delocalization of carriers are well described by either carrier-carrier interaction or disorder. When both effects come into play, however, a comprehensive understanding is not well established mainly due to complexity and sparse experimental data. Recently developed two-dimensional layered materials are ideal in describing such mesoscopic critical phenomena as they have both strong interactions and disorder. The transport in the insulating phase is well described by the soft Coulomb gap picture, which demonstrates the contribution of both interactions and disorder. Using this picture, we demonstrate the critical power law behavior of the localization length, supporting quantum criticality. We observe asymmetric critical exponents around the metal-insulator transition through temperature scaling analysis, which originates from poor screening in insulating regime and conversely strong screening in metallic regime due to free carriers. The effect of asymmetric scaling behavior is weakened in monolayer MoS2 due to a dominating disorder. © 2018 The Author(s
Activation Energies for the Fragmentation of Thiophene Ions by Surface-Induced Dissociation
Effects of ageing and arthritic disease on nitric oxide production by human articular chondrocytes
Energetics, timescales, and chemistry of low energy molecular ion-organic surface collisions
Protein Profiling of Mouse Livers with Peroxisome Proliferator-Activated Receptor α Activation
Peroxisome proliferator-activated receptor α (PPARα) is important in the induction of cell-specific pleiotropic responses, including the development of liver tumors, when it is chronically activated by structurally diverse synthetic ligands such as Wy-14,643 or by unmetabolized endogenous ligands resulting from the disruption of the gene encoding acyl coenzyme A (CoA) oxidase (AOX). Alterations in gene expression patterns in livers with PPARα activation were delineated by using a proteomic approach to analyze liver proteins of Wy-14,643-treated and AOX(−/−) mice. We identified 46 differentially expressed proteins in mouse livers with PPARα activation. Up-regulated proteins, including acetyl-CoA acetyltransferase, farnesyl pyrophosphate synthase, and carnitine O-octanoyltransferase, are involved in fatty acid metabolism, whereas down-regulated proteins, including ketohexokinase, formiminotransferase-cyclodeaminase, fructose-bisphosphatase aldolase B, sarcosine dehydrogenase, and cysteine sulfinic acid decarboxylase, are involved in carbohydrate and amino acid metabolism. Among stress response and xenobiotic metabolism proteins, selenium-binding protein 2 and catalase showed a dramatic ∼18-fold decrease in expression and a modest ∼6-fold increase in expression, respectively. In addition, glycine N-methyltransferase, pyrophosphate phosphohydrolase, and protein phosphatase 1D were down-regulated with PPARα activation. These observations establish proteomic profiles reflecting a common and predictable pattern of differential protein expression in livers with PPARα activation. We conclude that livers with PPARα activation are transcriptionally geared towards fatty acid combustion