SLCO/OATP-like transport of glutathione in FasL-induced apoptosis: glutathione efflux is coupled to an organic anion exchange and is necessary for the progression of the execution phase of apoptosis.
AuthorsFranco R,Cidlowski JA
JournalThe Journal of biological chemistry
PubMed ID16857677
DNA sequencing with dye-labeled terminators and T7 DNA polymerase: effect of dyes and dNTPs on incorporation of dye-terminators and probability analysis of termination fragments.
The incorporation of fluorescently labeled dideoxynucleotides by T7 DNA polymerase is optimized by the use of Mn2+, fluorescein analogs and four 2'-deoxyribonucleoside 5'-O-(1-thiotriphosphates) (dNTP alpha S's). The one-tube extension protocol was tested on single-stranded templates, as well as PCR fragments which were made single-stranded by digestion with T7 gene 6 ... More
Temperature dependence of membrane ion conductance analyzed by using the amphiphilic anion 5/6-carboxyfluorescein.
AuthorsBramhall J, Hofmann J, DeGuzman R, Montestruque S, Schell R
JournalBiochemistry
PubMed ID3427008
'The rate of efflux of trapped 5/6-carboxyfluorescein from sealed lipid vesicles showed a marked dependence on (a) temperature, (b) phospholipid acyl chain composition, and (c) the nature of co-trapped counterions. When the dye was salted with sodium, at pH greater than 7, the rate of dye permeation showed a discrete ... More
Structural transitions associated with the interaction of Alzheimer beta-amyloid peptides with gangliosides.
AuthorsMcLaurin J, Franklin T, Fraser PE, Chakrabartty A
JournalJ Biol Chem
PubMed ID9468505
'Alzheimer''s disease is characterized pathologically by the presence of neurofibrillary tangles and amyloid plaques. The principal component of the plaque is the beta-amyloid peptide (Abeta), a 39-43-residue peptide. The conformational change required for the conversion of soluble peptide into amyloid fibrils is modulated by pH, Abeta concentration, addition of kinetic ... More
The fate of Pluronic F-68 in chondrocytes and CHO cells.
AuthorsGigout A, Buschmann MD, Jolicoeur M,
JournalBiotechnol Bioeng
PubMed ID18393312
'The surfactant Pluronic F-68 (PF-68) is widely used in large-scale mammalian cell culture to protect cells from shear stress that arises from agitation and gas sparging. Several studies suggested that PF-68 is incorporated into the cell plasma membrane and could enter the cells, but without providing any direct evidence. The ... More
Comparative photostability studies of BODIPY and fluorescein dyes by using fluorescence correlation spectroscopy.
AuthorsHinkeldey B, Schmitt A, Jung G,
JournalChemphyschem
PubMed ID18816535
In single-molecule applications, the photostability of fluorescent molecules is a key parameter. We apply fluorescence correlation spectroscopy to compare the photostability of four fluorescein and four borondipyrromethene (BODIPY) dyes of similar structure but different triplet yields. The latter class of dyes are more stable. In the kinetic analysis the, diffusion ... More
Membrane disruption by Alzheimer beta-amyloid peptides mediated through specific binding to either phospholipids or gangliosides. Implications for neurotoxicity.
AuthorsMcLaurin J, Chakrabartty A
JournalJ Biol Chem
PubMed ID8900116
Increasing evidence implicates Abeta peptides as neurotoxic agents in Alzheimer's disease. We investigated one possible mechanism of neurotoxicity, namely Abeta-membrane lipid interactions. We find that Abeta disrupts membranes containing acidic phospholipids. This disruption is greater at slightly acidic pH (characteristic of endosomes) than at neutral pH (characteristic of the extracellular ... More
Connexins regulate calcium signaling by controlling ATP release.
AuthorsCotrina ML, Lin JH, Alves-Rodrigues A, Liu S, Li J, Azmi-Ghadimi H, Kang J, Naus CC, Nedergaard M
JournalProc Natl Acad Sci U S A
PubMed ID9861039
Forced expression of gap junction proteins, connexins, enables gap junction-deficient cell lines to propagate intercellular calcium waves. Here, we show that ATP secretion from the poorly coupled cell lines, C6 glioma, HeLa, and U373 glioblastoma, is potentiated 5- to 15-fold by connexin expression. ATP release required purinergic receptor-activated intracellular Ca2+ ... More
Characterization of 5(6)-carboxy-2,'7'-dichlorofluorescein transport by MRP2 and utilization of this substrate as a fluorescent surrogate for LTC4.
AuthorsHeredi-Szabo K, Kis E, Molnar E, Gyorfi A, Krajcsi P,
JournalJ Biomol Screen
PubMed ID18349419
MRP2 (ABCC2) is an efflux transporter expressed on the apical membrane of polarized cells. This protein has a major role in the biliary elimination of toxic compounds from the liver. As MRP2 transports many endogenous compounds, including LTC4 as well as xenobiotics and toxic phase II metabolites, blockade of this ... More
Advantages and limitations of commonly used methods to assay the molecular permeability of gap junctional intercellular communication.
AuthorsAbbaci M, Barberi-Heyob M, Blondel W, Guillemin F, Didelon J,
JournalBiotechniques
PubMed ID18611167
The role of gap junctional intercellular communication (GJIC) in regulation of normal growth and differentiation is becoming increasingly recognized as a major cellular function. GJIC consists of intercellular exchange of low molecular weight molecules, and is the only means for direct contact between cytoplasms of adjacent animal cells. Disturbances of ... More
Xanthene-dye-labelled phosphatidylethanolamines as probes of interfacial pH. Studies in phospholipid vesicles.
AuthorsKnight CG, Stephens T
JournalBiochem J
PubMed ID2471509
We have been developing the use of plasma-membrane-bound fluorescent probes to measure the pH values at the surfaces of living chondrocytes. For this purpose, three lipophilic pH indicators were made by covalently binding the xanthene dyes fluorescein, eosin or dichlorofluorescein to the amino group of phosphatidylethanolamine. The probes were incorporated ... More
Dicarboxy-dichlorofluorescein: a new fluorescent probe for measuring acidic intracellular pH.
AuthorsNedergaard M, Desai S, Pulsinelli W
JournalAnal Biochem
PubMed ID2372106
Derivatives of fluorescein sensitive to pH are extensively utilized for the determination of intracellular pH (pHi). Available dyes have pKa values of approximately 7.0, and are not well suited for measuring acidic pHi. We examined the fluorescein derivative, 5 (and 6)-carboxy-2',7'-dichlorofluorescein (CDCF) for its potential in the microspectrofluorometric measurement of ... More
Characterization of visible dyes for four-decay fluorescence detection in DNA sequencing.
AuthorsNunnally BK, He H, Li LC, Tucker SA, McGown LB
JournalAnal Chem
PubMed ID9212706
Dyes of several classes were investigated as candidates for use in a multiplex, four-decay fluorescence detection scheme for DNA sequencing. The dyes include nitrobenzofuran dyes, rhodamine dyes, fluorescein dyes, cyanine dyes, Nile Red, and BODIPY dyes. Based on the results of fluorescence spectral and lifetime studies, an initial set of ... More
Use of the anionic dye carboxy-2',7'-dichlorofluorescein for sensitive flow cytometric detection of multidrug resistance-associated protein activity.
AuthorsLaupeze B, Amiot L, Courtois A, Vernhet L, Drenou B, Fauchet R, Fardel O
JournalInt J Oncol
PubMed ID10427142
Multidrug resistance-associated protein (MRP) and P-glycoprotein are drug efflux pumps conferring multidrug resistance to tumor cells and sharing numerous substrates. In order to determine a flow cytometric assay allowing to analyse MRP activity in cancerous cells in a sensitive and specific manner, cellular accumulation and efflux of the anionic fluorescent ... More
Second messengers regulate endosomal acidification in Swiss 3T3 fibroblasts.
AuthorsZen K, Biwersi J, Periasamy N, Verkman AS
JournalJ Cell Biol
PubMed ID1382079
Acidification of the endosomal pathway is important for ligand and receptor sorting, toxin activation, and protein degradation by lysosomal acid hydrolases. Fluorescent probes and imaging methods were developed to measure pH to better than 0.2 U accuracy in individual endocytic vesicles in Swiss 3T3 fibroblasts. Endosomes were pulse labeled with ... More
Pharmacokinetics of 5 (and 6)-carboxy-2',7'-dichlorofluorescein and its diacetate promoiety in the liver.
Hepatic disposition of 5 (and 6)-carboxy-2',7'-dichlorofluorescein (CDF) and its diacetate promoiety (CDFDA) was studied in isolated perfused rat livers. Livers from Wistar wild-type and multidrug resistance-associated protein (Mrp)2-deficient (TR(-)) rats were perfused with CDF in the presence or absence of probenecid. Probenecid decreased the recovery of CDF in bile approximately ... More