Reactivity of mitochondrial sulfhydryl groups toward dithionitrobenzoic acid and bromobimanes under oligomycin-inhibited and uncoupling conditions.
AuthorsFreisleben HJ, Fuchs J, Mainka L, Zimmer G
JournalArch Biochem Biophys
PubMed ID2845867
Thiol reactivity was determined in rat heart mitochondria using chromophores of differing polarities: monobromobimane (MB), dithionitrobenzoate (NbS2), and bromobimane-q (MQ). The purpose of this study is to correlate reaction rates of protein thiols in the mitochondrial membrane with the oligomycin-inhibited and uncoupled states: In all cases investigated the reactivity of ... More
Thiol-specific probes indicate that the beta-chain of platelet glycoprotein Ib is a transmembrane protein with a reactive endofacial sulfhydryl group.
AuthorsKalomiris EL, Coller BS
JournalBiochemistry
PubMed ID2934089
We used two membrane-permeable fluorescent reagents, monobromobimane and N-[[5-(dimethylamino)-1-naphthalenyl]sulfonyl]aziridine (N-dansylaziridine), and one membrane-impermeable fluorescent probe, monobromo(trimethylammonio)bimane, all three of which react selectively with protein thiols, to assess the presence of reactive sulfhydryls in the platelet glycoprotein Ib (GPIb) molecule and establish the topology of any GPIb-reactive thiols in the platelet ... More
Accessibility of cell surface thiols in human lymphocytes is altered by ionophores or OKT-3 antibody.
AuthorsBalázs M, Matkó J, Szöllösi J, Mátyus L, Fulwyler MJ, Damjanovich S
JournalBiochem Biophys Res Commun
PubMed ID3675722
'The accessibility of cell surface sulfhydryl groups in human peripheral lymphocytes was investigated with 5,5''-dithiobis-(2-nitrobenzoic acid) in the presence and absence of ionophore antibiotics and the monoclonal antibody, OKT-3. Only a few accessible protein thiols have been found on the cells as demonstrated by labeling with a fluorescent non-penetrating thiol-marker, ... More
Bimane fluorescent labels. Characterization of the bimane labeling of human hemoglobin.
AuthorsKosower NS, Newton GL, Kosower EM, Ranney HM
JournalBiochim Biophys Acta
PubMed ID7378449
'The products of the bimane labeling (using a monobromobimane, a dibromobimane and a quaternary bromobimane) of hemoglobin are characterized. Peptide mapping identifies cysteine-beta 93 as the reactive thiol site. Electrophoretic mobility of hemoglobin varies with the label used, that of monobromobimane-labeled hemoglobin being unaltered, while dibromobimane- and trimethylammoniobromobimane-labeled hemoglobin exhibit ... More
Ligand-triggered conformational perturbations elicit changes at the single cysteinyl residue of spinach calmodulin.
AuthorsYuan SX, Haug A
JournalEur J Biochem
PubMed ID2841122
'Following application of stoichiometric amounts of Ca2+ or specific partner peptides to spinach calmodulin, dynamic changes in the nanosecond range could be monitored at a strategically anchored fluorescence or spin probe. For these studies the single cysteinyl residue 26 of spinach calmodulin was labelled with a thiol-specific proxyl (i.e. 2,2,5,5-tetramethyl-1-pyrrolidinyl-oxyl) ... More
Architecture of a K+ channel inner pore revealed by stoichiometric covalent modification.
AuthorsLu T, Nguyen B, Zhang X, Yang J
JournalNeuron
PubMed ID10197536
'Inwardly rectifying K+ channels bind intracellular magnesium and polyamines to generate inward rectification. We have examined the architecture of the inner pore of Kir2.1 channels by covalently attaching a constrained number (from one to four) of positively charged moieties of different sizes to the channel. Our results indicate that the ... More
Influence of pH on sulfhydryl groups and fluidity of the mitochondrial membrane.
AuthorsZimmer G, Freisleben HJ, Fuchs J
JournalArch Biochem Biophys
PubMed ID2173480
'Fluidity of the red blood cell membrane decreases as pH changes from 8 to 7.5. In rat liver mitochondrial (RLM) membrane fluidity precipitously declines as pH drops from 7.35 toward 7.0. With dithionitrobenzoate (Nbs2), reaction rates of mitochondrial -SH groups from rat liver and heart (RHM) and in beef heart ... More
Modification of platelet functions by monobromobimane, a fluorescent thiol group label.
AuthorsZucker MB, Mauss EA
JournalThromb Haemost
PubMed ID3012818
'Monobromobimane (mBBr, bimane), a compound that penetrates cells and forms a fluorescent adduct with thiol groups, was used to asses the significance of thiols in platelet function. Exposure of washed platelets for 1 min to 100 microM mBBr abolished ADP-induced aggregation; shape change was not inhibited by 500 microM mBBr. ... More
Effect of sulfhydryl group modification on the activity of bovine ferrochelatase.
AuthorsDailey HA
JournalJ Biol Chem
PubMed ID6698990
'The role of sulfhydryl groups in the activity of the terminal enzyme of the heme biosynthetic pathway, ferrochelatase (protoheme ferrolyase, EC 4.99.1.1), has been examined by using a variety of sulfhydryl group-specific reagents. The enzyme is rapidly inactivated in a pseudo-first order reaction by N-ethylmaleimide and monobromobimane and more slowly ... More
Analysis of biological thiols: derivatization with monobromotrimethylammoniobimane and characterization by electrophoresis and chromatography.
AuthorsFahey RC, Newton GL, Dorian R, Kosower EM
JournalAnal Biochem
PubMed ID6449160
Characterization of tubulin modified with the sulfhydryl-reactive fluorochrome monobromo(trimethylammonio)bimane.
AuthorsWadsworth P, Sloboda RD
JournalBiochemistry
PubMed ID7059577
Analysis of biological thiols: quantitative determination of thiols at the picomole level based upon derivatization with monobromobimanes and separation by cation-exchange chromatography.
AuthorsFahey RC, Newton GL, Dorian R, Kosower EM
JournalAnal Biochem
PubMed ID7247030
Bromobimane probes for thiols.
AuthorsKosower EM, Kosower NS
JournalMethods Enzymol
PubMed ID7651193
Thiol labeling with bromobimanes.
AuthorsKosower NS, Kosower EM
JournalMethods Enzymol
PubMed ID3657564
Bimane fluorescent labels: labeling of normal human red cells under physiological conditions.
AuthorsKosower NS, Kosower EM, Newton GL, Ranney HM
JournalProc Natl Acad Sci U S A
PubMed ID291011
The bimane fluorescent labels, monobromobimane, dibromobimane, and monobromotrimethylammoniobimane, are derivatives of syn-9,10-dioxabimane:1,5-diazabicyclo[3.3.0]octa-3,6-diene-2,8-dione. They efficiently label hemoglobin (reactive thiol groups), membrane proteins, and glutathione of normal human red cells under physiological conditions. Monobromobimane and dibromobimane are effective on intact cells while red cell membranes may be impermeable to the positively charged ... More
Distance measurement by energy transfer: the 3' end of 16-S RNA and proteins S4 and S17 of the ribosome of Escherichia coli.
Escherichia coli ribosomal proteins S4 and S17 were specifically labelled at their thiol groups with the acetylaminoethyl-dansyl and/or bimane fluorophores. Each formed a complex with 16-S RNA and, when the other 30-S ribosomal proteins were added, a complete 30-S subunit with at least partial activity. If the 3' end of ... More
Glutathione suppresses spontaneous activity in the frog spinal cord.
AuthorsLoguercio C, Di Cosmo A, De Santis A, Nardi G
JournalNeuroreport
PubMed ID8527738
Spontaneous electrical activity in the isolated hemisected frog spinal cord increased in the presence of the SH reductant dithiothreitol (DTT) was reversibly suppressed by the oxidant 5-5'-dithio-bis-(2 nitrobenzoic acid) (DTNB) and was irreversibly suppressed by the sulphydryl modifying agents N-ethyl-maleimide (NEM) and monobromotrimethylammonio bimane. Glutathione (GSH), an important natural low ... More
Neutrophil membrane sulfhydryl groups are involved in stimulated neutrophil adherence to endothelium.
AuthorsSchwartz BR, Harlan JM
JournalJ Leukoc Biol
PubMed ID2564411
We investigated the role of membrane sulfhydryl groups in adherence of stimulated polymorphonuclear neutrophils to cultured endothelial cells. Treatment of neutrophils with p-chloromercuriphenyl sulfonate (PCMPS), a slowly penetrating sulfhydryl reagent, inhibited phorbol myristate acetate (PMA)- or calcium ionophore A23187-stimulated adherence to cultured human or bovine endothelial cells. At concentrations which ... More
Nonequivalence of the nucleotide-binding subunits of an ABC transporter, the histidine permease, and conformational changes in the membrane complex.
AuthorsKreimer DI, Chai KP, Ferro-Luzzi Ames G
JournalBiochemistry
PubMed ID11087367
The membrane-bound complex of the Salmonella typhimurium histidine permease, an ABC transporter (or traffic ATPase), is composed of two membrane proteins, HisQ and HisM, and two identical copies of an ATP-hydrolyzing protein, HisP. We have developed a technique that monitors quantitatively the sulfhydryl modification levels within the intact complex, and ... More
Bromobimanes--fluorescent labeling agents for histochemical detection of sulfur containing neuropeptides in semithin sections.
AuthorsDanielsohn P, Nolte A
JournalHistochemistry
PubMed ID3570877
Sulfur containing neuropeptides could be demonstrated in semithin sections of invertebrate nervous tissue, especially of gastropods, by using the bromobimanes as fluorescent labeling agents for thiol groups. Semithin sections showed a brilliant fluorescence of labeled peptides and should be used if an excellent resolution is important. The three bromobimanes (MB: ... More
Bimane- and acrylodan-labeled S100 proteins. Role of cysteines-85 alpha and -84 beta in the conformation and calcium binding properties of S100 alpha alpha and S100b (beta beta) proteins.
AuthorsBaudier J, Glasser N, Duportail G
JournalBiochemistry
PubMed ID3801403
Bovine brain S100 alpha alpha, S100a (alpha beta), and S100b (beta beta) protein dimers were labeled with the sulfydryl-specific fluorescent probes monobromo(trimethylammonio)bimane (bimane) and 6-acryloyl-2-(dimethylamino)naphthalene (acrylodan) at cysteines-85 alpha and -84 beta. The conformation and fluorescence properties of the S100 proteins derived were studied by means of anion-exchange chromatography on ... More
A novel mycothiol-dependent detoxification pathway in mycobacteria involving mycothiol S-conjugate amidase.
AuthorsNewton GL, Av-Gay Y, Fahey RC
JournalBiochemistry
PubMed ID10978158
Mycothiol, 1-D-myo-inosityl-2-(N-acetylcysteinyl)amido-2-deoxy-alpha-D-glucopyranoside (MSH), is composed of N-acetylcysteine (AcCys) amide linked to 1-D-myo-inosityl-2-amino-2-deoxy-alpha-D-glucopyranoside (GlcN-Ins) and is the major thiol produced by most actinomycetes. When Mycobacterium smegmatis was treated with the alkylating agent monobromobimane (mBBr), the cellular mycothiol was converted to its bimane derivative (MSmB). The latter was rapidly cleaved to produce ... More
The "regulatory" sulfhydryl group of Penicillium chrysogenum ATP sulfurylase. Cooperative ligand binding after SH modification; chemical and thermodynamic properties.
ATP sulfurylase from Penicillium chrysogenum is a homohexamer that contains three free sulfhydryl groups/subunit, only one of which (designated SH-1) can be modified by disulfide, maleimide, and halide reagents under nondenaturing conditions. Modification of SH-1 has only a small effect on kcat but causes the [S]0.5 values for MgATP and ... More
Quantitative in vivo measurement of glutathione in Arabidopsis cells.
AuthorsMeyer AJ, May MJ, Fricker M
JournalPlant J
PubMed ID11489184
A new, non-destructive assay is described to quantify cytoplasmic glutathione (GSH) levels in vivo in single cells or populations of cells from Arabidopsis suspension cultures. Cytoplasmic GSH was labelled with monochlorobimane (MCB) in situ to give a fluorescent GSH-bimane (GSB) conjugate. At low (10-100 microM) concentrations of MCB, labelling was ... More
Fluorescent localization of thiols and disulfides in marsupial spermatozoa by bromobimane labelling.
AuthorsMate KE, Kosower NS, White IG, Rodger JC
JournalMol Reprod Dev
PubMed ID8185937
The acrosome of marsupial spermatozoa is a robust structure which, unlike its placental counterpart, resists disruption by detergent or freeze/thawing and does not undergo a calcium ionophore induced acrosome reaction. In this study specific fluorescent thiol labels, bromobimanes, were used to detect reactive thiols in the intact marsupial spermatozoon and ... More
Binding to phospholipid vesicles impairs substrate-mediated conformational changes of the precursor to mitochondrial aspartate aminotransferase.
AuthorsBerezov A, Iriarte A, Martinez-Carrion M
JournalJ Biol Chem
PubMed ID8071348
Specific labeling of both the mature (mAspAT) and precursor (pmAspAT) forms of rat liver mitochondrial aspartate aminotransferase with three different spectroscopic probes (monobromotrimethylammoniobimane, N-(iodoacetylaminoethyl)-5-naphthalene-1-sulfonic acid, and N-(1-pyrenyl)maleimide) was used to assess the possible conformational consequences of the interaction of a mitochondrial precursor protein with lipid membranes by means of fluorescence ... More