Cytokine-induced activation of nuclear factor-kappa B is inhibited by hydrogen peroxide through oxidative inactivation of IkappaB kinase.
AuthorsKorn SH, Wouters EF, Vos N, Janssen-Heininger YM
JournalJ Biol Chem
PubMed ID11479295
'Rapid activation of the IkappaB kinase (IKK) complex is considered an obligatory step in the activation of nuclear factor-kappaB (NF-kappaB) in response to diverse stimuli. Since oxidants have been implicated in the regulation of NF-kappaB, the focus of the present study was the activation of IKK by tumor necrosis factor ... More
Identification of proteins containing cysteine residues that are sensitive to oxidation by hydrogen peroxide at neutral pH.
AuthorsKim JR, Yoon HW, Kwon KS, Lee SR, Rhee SG
JournalAnal Biochem
PubMed ID10906242
'A procedure for detecting proteins that contain H(2)O(2)-sensitive cysteine (or selenocysteine) residues was developed as a means with which to study protein oxidation by H(2)O(2) in cells. The procedure is based on the facts that H(2)O(2) and biotin-conjugated iodoacetamide (BIAM) selectively and competitively react with cysteine residues that exhibit a ... More
In vivo dynamics of myosin II in Dictyostelium by fluorescent analogue cytochemistry.
AuthorsChu Q, Fukui Y
JournalCell Motil Cytoskeleton
PubMed ID8913645
'We used fluorescent analogue cytochemistry to study in vivo dynamics of myosin II in Dictyostelium discoideum. We labeled myosin with biotin or tetramethyl-rhodamine iodoacetamide (IATR). The labeled myosin shows normal activities as reversible filament assembly and Ca2+ and actin-activatable Mg(2+)-ATPase. We used the biotin-myosin as a probe examining the effects ... More
Molecular cloning and characterization of a mitochondrial selenocysteine-containing thioredoxin reductase from rat liver.
AuthorsLee SR, Kim JR, Kwon KS, Yoon HW, Levine RL, Ginsburg A, Rhee SG
JournalJ Biol Chem
PubMed ID9988709
'A thioredoxin reductase (TrxR), named here TrxR2, that did not react with antibodies to the previously identified TrxR (now named TrxR1) was purified from rat liver. Like TrxR1, TrxR2 was a dimeric enzyme containing selenocysteine (Secys) as the COOH-terminal penultimate residue. A cDNA encoding TrxR2 was cloned from rat liver; ... More
S-glutathiolation by peroxynitrite of p21ras at cysteine-118 mediates its direct activation and downstream signaling in endothelial cells.
AuthorsClavreul N, Adachi T, Pimental DR, Ido Y, Schöneich C, Cohen RA
JournalFASEB J
PubMed ID16415107
'The highly reactive species, peroxynitrite, is produced in endothelial cells in pathological states in which the production of superoxide anion and NO is increased. Here, we show that peroxynitrite added exogenously or generated endogenously in response to exposure to an NO donor or oxidized low-density lipoproteins (oxLDL) increases p21ras activity ... More
Purification and characterization of membrane proteins.
AuthorsKraehenbuhl JP, Bonnard C
JournalMethods Enzymol
PubMed ID2167430
In vitro evolution of a self-alkylating ribozyme.
AuthorsWilson C, Szostak JW
JournalNature
PubMed ID7723823
'RNA enzymes are postulated to have catalysed all chemical reactions in the earliest living cells. This idea is now investigated in a search for alkyl transferases from a pool of random sequence RNAs. Selection for self-biotinylation yields a transfer RNA-like ribozyme that efficiently catalyses carbon-nitrogen bond formation. Ribozymes can thus ... More
Methods for determining the modification of protein thiols by reactive lipids.
AuthorsOh J, Johnson MS, Landar A,
JournalMethods Cell Biol
PubMed ID17445707
Biotin-based tagging techniques have been applied with success to monitor the posttranslational modification of reactive protein thiols in cell signaling proteins and enzymes by ROS/RNS and electrophilic compounds. One of the most versatile techniques is to attach biotin covalently to thiol-reactive molecules, and label thiol-containing proteins. After separation ... More
Photoaffinity labeling of antibodies for applications in homogeneous fluoroimmunoassays.
AuthorsChang IN, Lin JN, Andrade JD, Herron JN
JournalAnal Chem
PubMed ID7762830
A homogeneous noncompetitive immunoassay based on photoaffinity labeling techniques is described. Using this method, a fluorophore (reporter) can be specifically attached to an antibody in the vicinity of its antigen-combining sites. Upon antigen binding, changes in the fluorescence spectrum of the reporter molecule are often observed. Two fluorophores, pyrene and ... More
Enteric commensal bacteria potentiate epithelial restitution via reactive oxygen species-mediated inactivation of focal adhesion kinase phosphatases.
AuthorsSwanson PA, Kumar A, Samarin S, Vijay-Kumar M, Kundu K, Murthy N, Hansen J, Nusrat A, Neish AS,
JournalProc Natl Acad Sci U S A
PubMed ID21555563
The mechanisms by which enteric commensal microbiota influence maturation and repair of the epithelial barrier are relatively unknown. Epithelial restitution requires active cell migration, a process dependent on dynamic turnover of focal cell-matrix adhesions (FAs). Here, we demonstrate that natural, commensal bacteria stimulate generation of reactive oxygen species (ROS) in ... More
S-glutathionylation of the Rpn2 regulatory subunit inhibits 26 S proteasomal function.
AuthorsZmijewski JW, Banerjee S, Abraham E,
JournalJ Biol Chem
PubMed ID19549781
Although increased intracellular concentrations of hydrogen peroxide (H2O2) are associated with inhibition of 26 S proteasomal activity, the mechanisms responsible for such effects have not been well delineated. In the present studies, we found that direct exposure of purified 26 S proteasomes to H2O2 had negligible effects on their activity, ... More
Selective enrichment of thiophosphorylated polypeptides as a tool for the analysis of protein phosphorylation.
AuthorsKwon SW, Kim SC, Jaunbergs J, Falck JR, Zhao Y,
JournalMol Cell Proteomics
PubMed ID12734387
A chemoselective alkylation method is described for the isolation and subsequent identification of thiophosphorylated peptides/proteins. The method involves thiophosphorylation of proteins using adenosine 5'-O-(thiotriphosphate) (ATPgammaS) followed by selective in situ alkylation of the newly thiophosphorylated proteins resulting in a stable covalent bond. The chemoselective alkylation exploits the relatively high nucleophilicity ... More
Modulation of cell surface protein free thiols: a potential novel mechanism of action of the sesquiterpene lactone parthenolide.
AuthorsSkalska J, Brookes PS, Nadtochiy SM, Hilchey SP, Jordan CT, Guzman ML, Maggirwar SB, Briehl MM, Bernstein SH,
JournalPLoS One
PubMed ID19956548
BACKGROUND: There has been much interest in targeting intracellular redox pathways as a therapeutic approach for cancer. Given recent data to suggest that the redox status of extracellular protein thiol groups (i.e. exofacial thiols) effects cell behavior, we hypothesized that redox active anti-cancer agents would modulate exofacial protein thiols. METHODOLOGY/PRINCIPAL ... More
Biotinylation reagents for the study of cell surface proteins.
AuthorsElia G,
JournalProteomics
PubMed ID18763706
The extraordinarily stable, non-covalent interaction between avidin and biotin is one of the most commonly exploited tools in chemistry and biology. Methods for derivatization with biotin of a variety of molecules (in particular, proteins) have been introduced, in order to allow their efficient recovery, immobilization and detection with avidin-based reagents. ... More
Reversibility of covalent electrophile-protein adducts and chemical toxicity.
AuthorsLin D, Saleh S, Liebler DC,
JournalChem Res Toxicol
PubMed ID19548357
The biotin-tagged electrophiles 1-biotinamido-4-(4'-[maleimidoethylcyclohexane]-carboxamido)butane (BMCC) and N-iodoacetyl-N-biotinylhexylenediamine (IAB) have been used as model electrophile probes in complex proteomes to identify protein targets associated with chemical toxicity. Whereas IAB activates stress signaling and apoptosis in HEK293 cells, BMCC does not. Cysteine Michael adducts formed from BMCC and nonbiotinylated analogues rapidly disappeared ... More
Redox regulation of beta-actin during integrin-mediated cell adhesion.
Redox sensitivity of actin toward an exogenous oxidative stress has recently been reported. We report here the first evidence of in vivo actin redox regulation by a physiological source of reactive oxygen species, specifically those species generated by integrin receptors during cell adhesion. Actin oxidation takes place via the formation ... More
The 'blue-on' opponent pathway in primate retina originates from a distinct bistratified ganglion cell type.
AuthorsDacey DM, Lee BB
JournalNature
PubMed ID8107868
Colour vision in humans and Old World monkeys begins with the differential activation of three types of cone photoreceptor which are maximally sensitive to short (S), medium (M) and long (L) wavelengths. Signals from the three cone types are relayed to the retinal ganglion cells via cone-specific bipolar cell types. ... More