Synthesis and properties of fluorescent beta-adrenoceptor ligands.
AuthorsHeithier H, Hallmann D, Boege F, Reiländer H, Dees C, Jaeggi KA, Arndt-Jovin D, Jovin TM, Helmreich EJ
JournalBiochemistry
PubMed ID7914090
We describe the synthesis of bordifluoropyrromethene (BODIPY), fluorescein, and related fluorescent derivatives of the beta-adrenergic ligand CGP 12177. With these probes we screened insect (Sf9) cells stably transformed with the human beta 2-adrenoceptor gene and expressing (2-3.5) x 10(5) human beta 2-adrenoceptors per cell. Among these derivatives only BODIPY-CGP gave ... More
LDL-mediated interaction of Lp[a] with HepG2 cells: a novel fluorescence microscopy approach.
AuthorsHofer G, Steyrer E, Kostner GM, Hermetter A
JournalJ Lipid Res
PubMed ID9458265
'We studied the topography of Lp[a]-LDL-cell interactions by means of fluorescence microscopy, using fluorescence-labeled lipoproteins. In contrast to known methods which are based on noncovalent labeling of lipoproteins by positively charged amphiphiles, the protein moiety of LDL and Lp[a] was covalently labeled with either BODIP-succinimide-ester (green) or rhodamine X iodoacetamide ... More
Interaction of a small heat shock protein of the fission yeast, Schizosaccharomyces pombe, with a denatured protein at elevated temperature.
AuthorsHirose M, Tohda H, Giga-Hama Y, Tsushima R, Zako T, Iizuka R, Pack C, Kinjo M, Ishii N, Yohda M
JournalJ Biol Chem
PubMed ID16055437
'We have expressed, purified, and characterized one small heat shock protein of the fission yeast Schizosaccharomyces pombe, SpHsp16.0. SpHsp16.0 was able to protect citrate synthase from thermal aggregation at 45 degrees C with high efficiency. It existed as a hexadecameric globular oligomer near the physiological growth temperature. At elevated temperatures, ... More
A comparison of the emission efficiency of four common green fluorescence dyes after internalization into cancer cells.
AuthorsHama Y, Urano Y, Koyama Y, Bernardo M, Choyke PL, Kobayashi H
JournalBioconjug Chem
PubMed ID17105220
'In vivo optical imaging to enhance the detection of cancer during endoscopy or surgery requires a targeted fluorescent probe with high emission efficiency and high signal-to-background ratio. One strategy to accurately detect cancers is to have the fluorophore internalize within the cancer cells permitting nonbound fluorophores to be washed away ... More
On-the-fly fluorescence lifetime detection of labeled DNA primers.
AuthorsLi LC, He H, Nunnally BK, McGown LB
JournalJ Chromatogr B Biomed Sci Appl
PubMed ID9271132
'The first application of frequency-domain, on-the-fly fluorescence lifetime detection to the detection, identification and resolution of fluorescent labeled oligonucleotide primers in capillary electrophoresis is presented. These studies, which are directed towards the development of four-decay DNA sequencing, employed two common sequencing primers, SP6 and M13/pUC. The primers were each tagged ... More
Gateable nanofluidic interconnects for multilayered microfluidic separation systems.
'The extension of microfluidic devices to include three-dimensional fluidic networks allows complex fluidic and chemical manipulations but requires innovative methods to interface fluidic layers. Externally controllable interconnects, employing nuclear track-etched polycarbonate membranes containing nanometer-diameter capillaries, are described that produce hybrid three-dimensional fluidic architectures. Controllable nanofluidic transfer is achieved by controlling ... More
Toxicity of organic fluorophores used in molecular imaging: literature review.
AuthorsAlford R, Simpson HM, Duberman J, Hill GC, Ogawa M, Regino C, Kobayashi H, Choyke PL,
JournalMol Imaging
PubMed ID20003892
'Fluorophores are potentially useful for in vivo cancer diagnosis. Using relatively inexpensive and portable equipment, optical imaging with fluorophores permits real-time detection of cancer. However, fluorophores can be toxic and must be investigated before they can be administered safely to patients. A review of published literature on the toxicity of ... More
Matrix-assisted laser desorption/ionization mass spectrometry of deoxynucleotides labeled with an IMI dye.
AuthorsLan ZH, Wang P, Giese RW
JournalRapid Commun Mass Spectrom
PubMed ID10407338
'The four major deoxynucleotides of DNA, and adduct mixtures resulting from separate reactions of 5''-dAMP and 5''-dGMP with benzo[a]pyrene diolepoxide (BPDE), were analyzed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) after labeling of their phosphate group with an IMI dye. The latter reagent comprises an imidazole functional group attached to ... More
Effects of gel material on fluorescence lifetime detection of dyes and dye-labeled DNA primers in capillary electrophoresis.
AuthorsLi L, McGown LB
JournalJ Chromatogr A
PubMed ID10360329
'Investigations of fluorescence lifetimes of the dye 6-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]hexanoic acid (NBD-HA) and of DNA M13 primers labeled with NBD-HA, Cy3, rhodamine green and 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene++ +-3-propionic acid (BODIPY-FL) dyes in polyacrylamide gels of various degrees of crosslinking and different crosslinkers, and in a cellulose sieving buffer with different organic modifiers, are described. ... More
High affinity C10-Oeq ester derivatives of ryanodine. Activator-selective agonists of the sarcoplasmic reticulum calcium release channel.
'The plant alkaloids ryanodine and dehydroryanodine are specific and potent modulators of the sarcoplasmic reticulum calcium release channel. In the present study, acidic, basic, and neutral side chains esters of these diterpene compounds were prepared and their pharmacologic activities were assessed. Binding affinities of the novel C10-Oeq ester derivatives for ... More
In vivo fluorescence measurement of Na(+) concentration in the pericryptal space of mouse descending colon.
AuthorsThiagarajah JR, Jayaraman S, Naftalin RJ, Verkman AS
JournalAm J Physiol Cell Physiol
PubMed ID11698248
'A method involving surgical exposure of the colonic mucosa, fluorescent dye addition, and confocal microscopy has been developed for monitoring colonic crypt function in vivo in mice. Na(+) concentration in the extracellular pericryptal space of descending colon was measured using a low-affinity Na(+)-sensitive fluorescent indicator consisting of an Na(+)-sensitive chromophore ... More
Lanthanide-binding tags as luminescent probes for studying protein interactions.
AuthorsSculimbrene BR, Imperiali B
JournalJ Am Chem Soc
PubMed ID16734490
'Herein, we report a method for studying protein-peptide interactions which exploits the luminescence properties of Tb(III). Lanthanide-binding tags (LBTs) are short peptide sequences comprising 15-20 naturally occurring amino acids that bind Tb(III) with high affinity. These genetically encodable luminescent tags are smaller in size than the Aequorea victoria fluorescent proteins ... More
Fluorescence resonance energy transfer to probe human M1 muscarinic receptor structure and drug binding properties.
AuthorsIlien B, Franchet C, Bernard P, Morisset S, Weill CO, Bourguignon JJ, Hibert M, Galzi JL
JournalJ Neurochem
PubMed ID12694403
'Human M1 muscarinic receptor chimeras were designed (i) to allow detection of their interaction with the fluorescent antagonist pirenzepine labelled with Bodipy [558/568], through fluorescence resonance energy transfer, (ii) to investigate the structure of the N-terminal extracellular moiety of the receptor and (iii) to set up a fluorescence-based assay to ... More
Wavelength-resolved fluorescence detection in capillary electrophoresis.
AuthorsTimperman AT, Khatib K, Sweedler JV
JournalAnal Chem
PubMed ID7864386
'A multichannel laser-induced fluorescence detector for capillary electrophoresis is described. The detection system combines yoctomole limits of detection with the simultaneous acquisition of entire fluorescence emission spectra. An Ar/Kr mixed-gas ion laser provides great flexibility in excitation wavelengths, and a holographic grating and charge-coupled device detector combination allows a 500-nm ... More
DNA sequencing by capillary electrophoresis with four-decay fluorescence detection.
AuthorsHe H, McGown LB
JournalAnal Chem
PubMed ID11140750
'A scheme for multiplex detection of dye-labeled DNA fragments in DNA sequencing is described in which on-the-fly, frequency-domain fluorescence lifetime detection is used to discriminate among the dye-labeled fragments of the four terminal bases in a single-lane CE separation. Two four-dye systems were evaluated, one excited at 488 nm and ... More
Membrane protein proteolysis assayed by fluorescence quenching: assay of O-sialoglycoprotein endopeptidase.
AuthorsJiang P, Mellors A
JournalAnal Biochem
PubMed ID9606137
'The assay of the O-sialoglycoprotein endopeptidase of Pasteurella haemolytica has previously used the cleavage of 125I-labeled glycophorin A, measured by SDS-PAGE, autoradiography, gel-slicing, and scintillation counting. A new assay is based on the increased fluorescence which results from proteolytic cleavage of a fluorescence-quenched micellar substrate, 4,4-difluor-5,7-dimethyl-4-bora-3 alpha, 4 alpha-diaza-s-indacene-3-propionic acid ... More
Preparation of an IMI dye (imidazole functional group) containing a 4-(N,N-dimethylaminosulfonyl)-2,1,3-benzoxadiazole fluorophore for labeling of phosphomonoesters.
AuthorsLan ZH, Qian X, Giese RW
JournalJ Chromatogr A
PubMed ID10070766
'We are studying dye-imidazole conjugates ("IMI dyes") as reagents for labeling phosphomonoesters such as nucleotides. Previously we have employed a BODIPY dye in our IMI reagents, and analyzed the labeled products by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) involving an argon ion laser. (The BODIPY fluorophore is a 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene). ... More
Incorporation of fluorescently labeled nonnatural amino acids into proteins in an E. coli in vitro translation system.
AuthorsHohsaka T, Abe R, Shiraga K, Sisido M
JournalNucleic Acids Res Suppl
PubMed ID14510485
'Various nonnatural amino acids has been incorporated into proteins by using four-base codons in an E. coli in vitro translation system. Here, design and synthesis of novel fluorescently labeled nonnatural amino acids and their incorporation into proteins were investigated. Transfer RNAs that contained a CCCG anticodon and were aminoacylated with ... More
Retrocyclin: a primate peptide that protects cells from infection by T- and M-tropic strains of HIV-1.
AuthorsCole AM, Hong T, Boo LM, Nguyen T, Zhao C, Bristol G, Zack JA, Waring AJ, Yang OO, Lehrer RI
JournalProc Natl Acad Sci U S A
PubMed ID11854483
'Human bone marrow expresses a pseudogene that encodes an antimicrobial peptide homologous to rhesus monkey circular minidefensins (delta-defensins). We prepared the putative ancestral human peptide by solid-phase synthesis and named it "retrocyclin." Retrocyclin did not cause direct inactivation of HIV-1, and its modest antibacterial properties resembled those of its rhesus ... More
Printing proteins as microarrays for high-throughput function determination.
AuthorsMacBeath G, Schreiber SL
JournalScience
PubMed ID10976071
'Systematic efforts are currently under way to construct defined sets of cloned genes for high-throughput expression and purification of recombinant proteins. To facilitate subsequent studies of protein function, we have developed miniaturized assays that accommodate extremely low sample volumes and enable the rapid, simultaneous processing of thousands of proteins. A ... More
Selective photoaffinity labeling of the inositol polyphosphate binding C2B domains of synaptotagmins.
AuthorsMehrotra B, Elliott JT, Chen J, Olszewski JD, Profit AA, Chaudhary A, Fukuda M, Mikoshiba K, Prestwich GD
JournalJ Biol Chem
PubMed ID9020139
'Synaptotagmin (Syt) II, a synaptic vesicle protein containing two copies of highly conserved protein kinase C homology regions known as the C2A and C2B domains, acts as a Ca2+ sensor and provides both phospholipid and inositol polyphosphate (IPn) recognition domains important in endo- and exocytosis. Four photoaffinity analogues of IP3, ... More
Activatable fluorescent molecular imaging of peritoneal metastases following pretargeting with a biotinylated monoclonal antibody.
AuthorsHama Y, Urano Y, Koyama Y, Choyke PL, Kobayashi H
JournalCancer Res
PubMed ID17440095
'Optical probes that yield high target-to-background ratios are necessary to detect microfoci of cancer that would otherwise escape detection with white light imaging. Target-specific activation of the optical signal at tumor foci is one mechanism by which high target and low background signal can be achieved. Here, we describe a ... More
FRET analysis of protein conformational change through position-specific incorporation of fluorescent amino acids.
AuthorsKajihara D, Abe R, Iijima I, Komiyama C, Sisido M, Hohsaka T
JournalNat Methods
PubMed ID17060916
'We designed and synthesized new, fluorescent, non-natural amino acids that emit fluorescence of wavelengths longer than 500 nm and are accepted by an Escherichia coli cell-free translation system. We synthesized p-aminophenylalanine derivatives linked with BODIPY fluorophores at the p-amino group and introduced them into streptavidin using the four-base codon CGGG ... More
Capillary electrophoretic determination of cathepsin D activity using Oregon Green-labeled hemoglobin.
AuthorsChu Q, Jones S, Zeece M
JournalElectrophoresis
PubMed ID10546832
'Cathepsin D is an aspartyl protease of lysosomal origin and functions in a variety of roles including protein turnover, catabolism of peptide hormones, antigen processing and presentation, and neoplastic disease. In breast cancer, the level of cathepsin D has been linked to metastasis and prognosis for survivability. Many of these ... More
In vitro and in vivo intrapulmonary distribution of fluorescently labeled surfactant.
AuthorsDiemel RV, Walch M, Haagsman HP, Putz G
JournalCrit Care Med
PubMed ID12006806
'OBJECTIVE: To determine the distribution of endotracheally administered surfactant at the alveolar level in an animal model of acute respiratory distress syndrome. DESIGN: Prospective, randomized animal study. SETTING: Research laboratory of a university hospital. SUBJECTS: Seventy-one male Sprague-Dawley rats, weighing 330-370 g. INTERVENTIONS: To measure surfactant distribution in vitro, a ... More
Multicolor in vitro translation.
AuthorsTraverso G, Diehl F, Hurst R, Shuber A, Whitney D, Johnson C, Levin B, Kinzler KW, Vogelstein B
JournalNat Biotechnol
PubMed ID12910244
'In vitro translation is a widely used tool for both analytical and preparative purposes. For analytical purposes, small amounts of proteins are synthesized and visualized by detection of labeled amino acids incorporated during translation. The original strategy of incorporating radioactively labeled amino acids, such as [35S]methionine or [14C]leucine, has been ... More
Function-based isolation of novel enzymes from a large library.
'Here we describe a high-throughput, quantitative method for the isolation of enzymes with novel substrate specificities from large libraries of protein variants. Protein variants are displayed on the surface of microorganisms and incubated with a synthetic substrate consisting of (1) a fluorescent dye (2) a positively charged moiety (3) the ... More
Flow cytometry-based biosensor for detection of multivalent proteins.
AuthorsSong X, Shi J, Swanson B
JournalAnal Biochem
PubMed ID10933853
'Microsphere-based flow cytometric detection of cholera toxin (CT) through distance-dependent fluorescence resonant energy transfer (FRET) has been developed. Simultaneous double-fluorescence changes induced by multivalent interactions between CT and fluorophore (both fluorescence donor and acceptor)-labeled ganglioside GM1 on a biomimetic membrane surface (supported bilayers of phospholipids) can be measured by a ... More
Evaluation of disulfide reduction during receptor-mediated endocytosis by using FRET imaging.
AuthorsYang J, Chen H, Vlahov IR, Cheng JX, Low PS
JournalProc Natl Acad Sci U S A
PubMed ID16950881
'Despite functional evidence for disulfide bond-reducing activity in endosomal compartments, the mechanistic details pertaining to such process (e.g., kinetics and sites of disulfide reduction) remain largely controversial. To address these questions directly, we have synthesized a previously uncharacterized fluorescent folate conjugate, folate-(BODIPY FL)-SS-rhodamine (folate-FRET), that changes fluorescence from red to ... More
Color-blind fluorescence detection for four-color DNA sequencing.
AuthorsLewis EK, Haaland WC, Nguyen F, Heller DA, Allen MJ, MacGregor RR, Berger CS, Willingham B, Burns LA, Scott GB, Kittrell C, Johnson BR, Curl RF, Metzker ML
JournalProc Natl Acad Sci U S A
PubMed ID15800037
'We present an approach called pulsed multiline excitation (PME) for measurements of multicomponent, fluorescence species and demonstrate its application in capillary electrophoresis for DNA sequencing. To fully demonstrate the advantages of PME, a fluorescent dye set has been developed whose absorption maxima span virtually the entire visible spectrum. Unlike emission ... More
Synthesis of fluorescent biochemical tools related to the 2-azetidinone class of cholesterol absorption inhibitors.
AuthorsBurnett DA, Caplen MA, Browne ME, Zhau H, Altmann SW, Davis HR, Clader JW
JournalBioorg Med Chem Lett
PubMed ID11814786
'Fluorescent analogues of the cholesterol absorption inhibitor (CAI), Sch 58235, have been designed and synthesized as single enantiomers. Biological testing reveals that they are potent CAIs and are suitable tools for the investigation of the azetidinone CAI mechanism of action (MOA).' ... More
Nuclear localization of pyrrole-imidazole polyamide-fluorescein conjugates in cell culture.
AuthorsBest TP, Edelson BS, Nickols NG, Dervan PB
JournalProc Natl Acad Sci U S A
PubMed ID14519850
'A series of hairpin pyrrole-imidazole polyamide-fluorescein conjugates were synthesized and assayed for cellular localization. Thirteen cell lines, representing 11 human cancers, one human transformed kidney cell line, and one murine leukemia cell line, were treated with 5 microM polyamide-fluorescein conjugates for 10-14 h, then imaged by confocal laser scanning microscopy. ... More
DNA sequence-specific polyamides alleviate transcription inhibition associated with long GAA.TTC repeats in Friedreich's ataxia.
AuthorsBurnett R, Melander C, Puckett JW, Son LS, Wells RD, Dervan PB, Gottesfeld JM
JournalProc Natl Acad Sci U S A
PubMed ID16857735
'The DNA abnormality found in 98% of Friedreich''s ataxia (FRDA) patients is the unstable hyperexpansion of a GAA.TTC triplet repeat in the first intron of the frataxin gene. Expanded GAA.TTC repeats result in decreased transcription and reduced levels of frataxin protein in affected individuals. Beta-alanine-linked pyrrole-imidazole polyamides bind GAA.TTC tracts ... More
Fluorescent assay for studying the substrate specificity of neuraminidase.
Facile conversion of aptamers into sensors using a 2'-ribose-linked fluorophore.
AuthorsMerino EJ, Weeks KM
JournalJ Am Chem Soc
PubMed ID16159247
Using BODIPY dye-primer chemistry in large-scale sequencing.
AuthorsMuzny DM, Metzker ML, Bouck J, Gorrell JH, Ding Y, Maxim E, Gibbs RA
JournalIEEE Eng Med Biol Mag
PubMed ID9824768
Liquid-phase synthesis of a pegylated adenosine-oligoarginine conjugate, cell-permeable inhibitor of cAMP-dependent protein kinase.
AuthorsViht K, Padari K, Raidaru G, Subbi J, Tammiste I, Pooga M, Uri A
JournalBioorg Med Chem Lett
PubMed ID12941328
An adenosine-oligoarginine conjugate (ARC) was assembled in a stepwise manner on a poly(ethylene glycol) carrier. The pegylated conjugate inhibited cAMP-dependent protein kinase with IC(50)=460 nM and the cellular uptake of its BODIPY FL derivative was demonstrated and compared to that of free ARC with fluorescence microscopy. ... More
Cellular uptake of N-methylpyrrole/N-methylimidazole polyamide-dye conjugates.
AuthorsBelitsky JM, Leslie SJ, Arora PS, Beerman TA, Dervan PB
JournalBioorg Med Chem
PubMed ID12150878
The cellular uptake and localization properties of DNA binding N-methylpyrrole/N-methylimidazole polyamide-dye conjugates in a variety of living cells have been examined by confocal laser scanning microscopy. With the exception of certain T-cell lines, polyamide-dye conjugates localize mainly in the cytoplasm and not in the nucleus. Reagents such as methanol typically ... More
Gangliosides GM1 and GD1b are not polarized in mature hippocampal neurons.
Analysis of the binding of cholera toxin to ganglioside GM1 in both living and fixed neurons, and comparison with the distribution of defined axonal and dendritic proteins, demonstrates that ganglioside GM1 is distributed in a non-polarized manner over the axonal and dendritic plasma membranes of mature, cultured hippocampal neurons. Likewise, ... More
Multiplex capillary denaturing high-performance liquid chromatography with laser-induced fluorescence detection.
AuthorsXiao W, Stern D, Jain M, Huber CG, Oefner PJ
JournalBiotechniques
PubMed ID11414227
Denaturing high-performance liquid chromatography (DHPLC) is a sensitive, robust, and operationally inexpensive method for the detection of single-base substitutions and small deletions and insertions. To increase sample throughout, we have developed a multiplexing strategy using fluorophores to distinguish different PCR products. The system combines recent advances in the synthesis of ... More
The global spread of malaria in a future, warmer world.
AuthorsRogers DJ, Randolph SE
JournalScience
PubMed ID10976072
The frequent warnings that global climate change will allow falciparum malaria to spread into northern latitudes, including Europe and large parts of the United States, are based on biological transmission models driven principally by temperature. These models were assessed for their value in predicting present, and therefore future, malaria distribution. ... More
Coupled transcription and translation within nuclei of mammalian cells.
AuthorsIborra FJ, Jackson DA, Cook PR
JournalScience
PubMed ID11423616
It is widely assumed that the vital processes of transcription and translation are spatially separated in eukaryotes and that no translation occurs in nuclei. We localized translation sites by incubating permeabilized mammalian cells with [3H]lysine or lysyl-transfer RNA tagged with biotin or BODIPY; although most nascent polypeptides were cytoplasmic, some ... More
Fluorescent quenching-based quantitative detection of specific DNA/RNA using a BODIPY((R)) FL-labeled probe or primer.
AuthorsKurata S, Kanagawa T, Yamada K, Torimura M, Yokomaku T, Kamagata Y, Kurane R
JournalNucleic Acids Res
PubMed ID11239011
We have developed a simple method for the quantitative detection of specific DNA or RNA molecules based on the finding that BODIPY((R)) FL fluorescence was quenched by its interaction with a uniquely positioned guanine. This approach makes use of an oligonucleotide probe or primer containing a BODIPY((R)) FL-modified cytosine at ... More
Detection of the mature, but not precursor, RNA using a fluorescent DNA probe.
AuthorsPaiboonskuwong K, Kato Y,
JournalNucleic Acids Symp Ser (Oxf)
PubMed ID17150950
Fluorescently labelled oligonucleotide probes have been widely used in biotechnology and diagnostic field. We improved the molecular beacon probe to detect the mature, but not precursor, RNA selectively. Based on the principle that a kind of fluorophore can be quenched by adjacent guanine base, therefore, the noise fluorescence caused by ... More
Electrophoretically uniform fluorescent dyes for automated DNA sequencing.
AuthorsMetzker ML, Lu J, Gibbs RA
JournalScience
PubMed ID8596915
A class of dyes, BODIPY fluorophores, has been identified for automated DNA sequencing that has improved spectral characteristics compared with conventional fluorescein and rhodamine dyes. Single and double BODIPY dye primers were characterized in commercially available DNA sequencers and showed uniform electrophoretic mobilities and high fluorescence intensities. The improved physical ... More
Controlling the intracellular localization of fluorescent polyamide analogues in cultured cells.
AuthorsCrowley KS, Phillion DP, Woodard SS, Schweitzer BA, Singh M, Shabany H, Burnette B, Hippenmeyer P, Heitmeier M, Bashkin JK
JournalBioorg Med Chem Lett
PubMed ID12699756
The intracellular distribution of fluorescent-labeled polyamides was examined in live cells. We showed that BODIPY-labeled polyamides accumulate in acidic vesicles, mainly lysosomes, in the cytoplasm of HCT116 colon cancer cells and human rheumatoid synovial fibroblasts (RSF). Verapamil blocked vesicular accumulation and led to nuclear accumulation of the BODIPY-labeled polyamide in ... More
Cell-free N-terminal protein labeling using initiator suppressor tRNA.
A highly efficient method for the introduction of fluorophores and other markers at the N terminus of proteins produced in a cell-free extract has been developed. The method utilizes an amber (CUA) initiator suppressor tRNA chemically aminoacylated with a fluorophore-amino acid conjugate which is introduced into an Escherichia coli S30 ... More
Solid-phase oligonucleotide synthesis and flow cytometric analysis with microspheres encoded with covalently attached fluorophores.
AuthorsNanthakumar A, Pon RT, Mazumder A, Yu S, Watson A
JournalBioconjug Chem
PubMed ID10725106
A novel combinatorial approach to synthesize oligonucleotides on fluorescently encoded microspheres based on flow sorting and segmental solid-phase synthesis is described. BODIPY dyes were covalently attached to polystyrene (8.8 microm, 55% DVB) microsphere particles to generate four fluorescently encoded sets. 20-mer oligonucleotide sequences can be synthesized on these microspheres with ... More
Chain dynamics of nascent polypeptides emerging from the ribosome.
Very little is known about the conformation of polypeptides emerging from the ribosome during protein biosynthesis. Here, we explore the dynamics of ribosome-bound nascent polypeptides and proteins in Escherichia coli by dynamic fluorescence depolarization and assess the population of cotranslationally active chaperones trigger factor (TF) and DnaK. E. coli cell-free ... More
Changes in retention behavior of fluorescently labeled proteins during ion-exchange chromatography caused by different protein surface labeling positions.
AuthorsTeske CA, Simon R, Niebisch A, Hubbuch J,
JournalBiotechnol Bioeng
PubMed ID17318908
Confocal laser scanning microscopy (CLSM) is a method allowing in situ visualization of protein transport in porous chromatography resins. CLSM requires labeling a protein with a fluorescent probe. Recent work has shown that conjugation of the protein with fluorescent probes can lead to significant changes in the retention time of ... More
Single-molecule imaging of a fluorescent unnatural amino acid incorporated into nicotinic receptors.
AuthorsPantoja R, Rodriguez EA, Dibas MI, Dougherty DA, Lester HA,
JournalBiophys J
PubMed ID19134478
We report on the first, to our knowledge, successful detection of a fluorescent unnatural amino acid (fUAA), Lys(BODIPYFL), incorporated into a membrane protein (the muscle nicotinic acetylcholine receptor, nAChR) in a living cell. Xenopus oocytes were injected with a frameshift-suppressor tRNA, amino-acylated with Lys(BODIPYFL) and nAChR (alpha/beta19'GGGU/gamma/delta) mRNAs. We measured ... More
NMR and bioinformatics discovery of exosites that tune metalloelastase specificity for solubilized elastin and collagen triple helices.
The catalytic domain of metalloelastase (matrix metalloproteinase-12 or MMP-12) is unique among MMPs in exerting high proteolytic activity upon fibrils that resist hydrolysis, especially elastin from lungs afflicted with chronic obstructive pulmonary disease or arteries with aneurysms. How does the MMP-12 catalytic domain achieve this specificity? NMR interface mapping suggests ... More
Import and fate of fluorescent analogs of oxidized phospholipids in vascular smooth muscle cells.
AuthorsMoumtzi A, Trenker M, Flicker K, Zenzmaier E, Saf R, Hermetter A,
JournalJ Lipid Res
PubMed ID17135656
Lipid oxidation is now thought to be an initiating and sustaining event in atherogenesis. Oxidatively fragmented phospholipids, namely 1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphocholine (PGPC) and 1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphocholine (POVPC), present in minimally modified LDL and atherosclerotic lesions, have been reported to elicit a wide range of pathophysiological responses in the cells of the vascular wall. Nevertheless, ... More
Four-color DNA sequencing with 3'-O-modified nucleotide reversible terminators and chemically cleavable fluorescent dideoxynucleotides.
AuthorsGuo J, Xu N, Li Z, Zhang S, Wu J, Kim DH, Sano Marma M, Meng Q, Cao H, Li X, Shi S, Yu L, Kalachikov S, Russo JJ, Turro NJ, Ju J,
JournalProc Natl Acad Sci U S A
PubMed ID18591653
DNA sequencing by synthesis (SBS) on a solid surface during polymerase reaction can decipher many sequences in parallel. We report here a DNA sequencing method that is a hybrid between the Sanger dideoxynucleotide terminating reaction and SBS. In this approach, four nucleotides, modified as reversible terminators by capping the 3'-OH ... More
Spectral fluorescence molecular imaging of lung metastases targeting HER2/neu.
AuthorsKoyama Y, Hama Y, Urano Y, Nguyen DM, Choyke PL, Kobayashi H,
JournalClin Cancer Res
PubMed ID17504994
PURPOSE: Surgical resection of pulmonary metastases is now a clinically accepted cancer therapy but its success depends on the accurate localization and removal of all tumor foci. To enhance the detection of pulmonary metastases during surgery, we developed an i.v. administered optical probe that uses a monoclonal antibody, Herceptin (trastuzumab), ... More
Influence of fluorophore and linker composition on the pharmacology of fluorescent adenosine A1 receptor ligands.
AuthorsBaker JG, Middleton R, Adams L, May LT, Briddon SJ, Kellam B, Hill SJ,
JournalBr J Pharmacol
PubMed ID20105183
BACKGROUND AND PURPOSE: The introduction of fluorescence-based techniques, and in particular the development of fluorescent ligands, has allowed the study of G protein-coupled receptor pharmacology at the single cell and single molecule level. This study evaluated how the physicochemical nature of the linker and the fluorophore affected the pharmacological properties ... More
Metabolic compartmentation in living cells: structural association of aldolase.
AuthorsWang J, Tolan DR, Pagliaro L
JournalExp Cell Res
PubMed ID9434640
The glycolytic enzyme aldolase is concentrated in a domain around stress fibers in living Swiss 3T3 cells, but the mechanism by which aldolase is localized has not been revealed. We have recently identified a molecular binding site for F-actin on aldolase, and we hypothesized that this specific binding interaction, rather ... More
Characterization of dye-induced mobility shifts affecting DNA sequencing in poly(ethylene oxide) sieving matrix.
AuthorsTan H, Yeung ES
JournalElectrophoresis
PubMed ID9504827
The influence of three classes of fluorescence labels including dipyrrometheneboron difluoride (BODIPY), energy transfer (ET) and conventional fluorescein and rhodamine (ABI), on DNA sequencing has been examined with laser-induced fluorescence detection and poly(ethylene oxide)-filled capillary electrophoresis. DNA sequencing fragments were generated by dye-labeled primer cycle-sequencing reactions in a hot-air thermal ... More
Isolation of primitive human hematopoietic progenitors on the basis of aldehyde dehydrogenase activity.
AuthorsStorms RW, Trujillo AP, Springer JB, Shah L, Colvin OM, Ludeman SM, Smith C
JournalProc Natl Acad Sci U S A
PubMed ID10430905
Because hematopoietic stem cells are rich in aldehyde dehydrogenase (ALDH) activity, we developed a fluorescent substrate for ALDH, termed BODIPY aminoacetaldehyde (BAAA), and tested its potential for isolating primitive human hematopoietic cells. A population of cells with low orthogonal light scattering and bright fluorescence intensity (SSC(lo)ALDH(br) cells) could be readily ... More
Comparison of fluorescence energy transfer primers with different donor-acceptor dye combinations.
AuthorsHung SC, Mathies RA, Glazer AN
JournalAnal Biochem
PubMed ID9448839
Fluorescence energy transfer (ET) primers are far superior to single dye-labeled primers as labels for DNA sequencing and polymerase chain reaction amplification. We compare here ET primers with different donor and acceptor dye combinations with respect to the relative acceptor fluorescence emission intensity and the amount of residual donor fluorescence ... More
Detection of hybrid formation between peptide nucleic acids and DNA by fluorescence polarization in the presence of polylysine.
AuthorsNikiforov TT, Jeong S
JournalAnal Biochem
PubMed ID10552912
A new method for the detection of PNA/DNA hybrids is presented. In this method, short PNA probes (9-13 mer) are labeled with a fluorescent dye and allowed to hybridize to target DNA molecules. A cationic polyamino acid, such as polylysine, is then added to the reaction mixture, whereupon the DNA ... More
A synthesis of 7 alpha-substituted estradiols: synthesis and biological evaluation of a 7 alpha-pentyl-substituted BODIPY fluorescent conjugate and a fluorine-18-labeled 7 alpha-pentylestradiol analog.
AuthorsFrench AN, Wilson SR, Welch MJ, Katzenellenbogen JA
JournalSteroids
PubMed ID8493705
In an effort to assist in the preparation of ligands for the study of the estrogen receptor (ER), we have developed a new synthesis of 7 alpha-substituted estradiols. The key step in the synthesis involves a copper-catalyzed, alpha-selective, 1,6-conjugate addition of 4-pentenyl magnesium bromide to a suitably protected 6-dehydrotestosterone derivative. ... More
Fluorescent verapamil derivative for monitoring activity of the multidrug transporter.
AuthorsLelong IH, Guzikowski AP, Haugland RP, Pastan I, Gottesman MM, Willingham MC
JournalMol Pharmacol
PubMed ID1681415
Multidrug resistance to amphipathic natural product chemotherapeutic drugs is conferred on cancer cells by expression of the MDR1 gene, which encodes the 170-kDa multidrug transporter known as P glycoprotein. The P glycoprotein-mediated efflux of toxic chemotherapeutic drugs can be reversed by agents such as verapamil, which is a substrate for ... More
Quantitation of mixed-base populations of HIV-1 variants by automated DNA sequencing with BODIPY dye-labeled primers.
AuthorsMetzker ML, Ansari-Lari MA, Liu XM, Holder DJ, Gibbs RA
JournalBiotechniques
PubMed ID9762443
Direct DNA sequencing of human immunodeficiency virus type 1 (HIV-1) pol and env gene regions was characterized for accuracy and precision. Restricted maximum likelihood (REML) analysis of molecular clone reconstruction experiments using a primer-walking strategy showed that the BODIPY and BODIPY energy-transfer (BET) dye primers sets were significantly more accurate ... More
Cholera toxin is found in detergent-insoluble rafts/domains at the cell surface of hippocampal neurons but is internalized via a raft-independent mechanism.
AuthorsShogomori H, Futerman AH
JournalJ Biol Chem
PubMed ID11113142
A number of studies have demonstrated that cholera toxin (CT) is found in detergent-insoluble, cholesterol-enriched domains (rafts) in various cells, including neurons. We now demonstrate that even though CT is associated with these domains at the cell surface of cultured hippocampal neurons, it is internalized via a raft-independent mechanism, at ... More
Visualizing association of N-ras in lipid microdomains: influence of domain structure and interfacial adsorption.
AuthorsNicolini C, Baranski J, Schlummer S, Palomo J, Lumbierres-Burgues M, Kahms M, Kuhlmann J, Sanchez S, Gratton E, Waldmann H, Winter R
JournalJ Am Chem Soc
PubMed ID16390147
In this study, two-photon fluorescence microscopy on giant unilamellar vesicles and tapping-mode atomic force microscopy (AFM) are applied to follow the insertion of a fluorescently (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene, BODIPY) labeled and completely lipidated (hexadecylated and farnesylated) N-Ras protein into heterogeneous lipid bilayer systems. The bilayers consist of the canonical raft mixture 1-palmitoyl-2-oleoylphosphatidylcholine ... More
Neutralization and transfer of lipopolysaccharide by phospholipid transfer protein.
AuthorsHailman E, Albers JJ, Wolfbauer G, Tu AY, Wright SD
JournalJ Biol Chem
PubMed ID8647810
Phospholipid transfer protein (PLTP) and lipopolysaccharide-binding protein (LPB) are lipid transfer proteins found in human plasma. PLTP shares 24% sequence similarity with LBP. PLTP mediates the transfer and exchange of phospholipids between lipoprotein particles, whereas LBP transfers bacterial lipopolysaccharide (LPS) either to lipoprotein particles or to CD14, a soluble and ... 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
Scanning two-photon fluctuation correlation spectroscopy: particle counting measurements for detection of molecular aggregation.
AuthorsBerland KM, So PT, Chen Y, Mantulin WW, Gratton E
JournalBiophys J
PubMed ID8804624
Scanning fluctuation correlation spectroscopy (FCS) is an experimental technique capable of measuring particle number concentrations by monitoring spontaneous equilibrium fluctuations in the local concentration of a fluorescent species in a small (femtoliter) subvolume of a sample. The method can be used to detect molecular aggregation for dilute, submicromolar samples by ... More
Receptor-induced internalization of selective peptidic mu and delta opioid ligands.
AuthorsGaudriault G, Nouel D, Dal Farra C, Beaudet A, Vincent JP
JournalJ Biol Chem
PubMed ID9006932
The binding and internalization of radioiodinated and fluorescent mu and delta opioid peptides in mammalian cells were quantitatively studied by biochemical techniques and directly visualized by confocal microscopy. The labeled peptides were prepared by inserting either a 125I-Bolton-Hunter group or a fluorescent probe into the C-terminal part of 5-aminopentylamide derivatives ... More
2,2'-thiodiethanol: a new water soluble mounting medium for high resolution optical microscopy.
AuthorsStaudt T, Lang MC, Medda R, Engelhardt J, Hell SW
JournalMicrosc Res Tech
PubMed ID17131355
The use of high numerical aperture immersion lenses in optical microscopy is compromised by spherical aberrations induced by the refractive index mismatch between the immersion system and the embedding medium of the sample. Especially when imaging >10 micro m deep into the specimen, the refractive index mismatch results in a ... More
Receptor-mediated targeting of fluorescent probes in living cells.
AuthorsFarinas J, Verkman AS
JournalJ Biol Chem
PubMed ID10075643
A strategy was developed to label specified sites in living cells with a wide selection of fluorescent or other probes and applied to study pH regulation in Golgi. cDNA transfection was used to target a single-chain antibody to a specified site such as an organelle lumen. The targeted antibody functioned ... More
Photodestruction of fluorophores and optimum conditions for trace DNA detection by automated DNA sequencer.
AuthorsKambara H, Nagai K, Kawamoto K
JournalElectrophoresis
PubMed ID1451690
Although automated DNA sequencers are becoming popular, their sensitivity in detecting DNA bands is still around 10(-17) mole/band. The sensitivity of a system depends on the laser power, labeling fluorophore, and the fluorescence-collecting yield. The emission and photodestruction cross-sections of the fluorophores are critical in optimizing the irradiated laser power ... More
Synthesis and characterization of 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene (BODIPY)-labeled fluorescent ligands for the mu opioid receptor.
AuthorsEmmerson PJ, Archer S, El-Hamouly W, Mansour A, Akil H, Medzihradsky F
JournalBiochem Pharmacol
PubMed ID9393674
A series of opioid ligands utilizing the 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene (BODIPY) fluorophores 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene++ +-3-propionic acid or 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza- s-indacene-3-propionic acid were synthesized and characterized for their ability to act as a suitable fluorescent label for the mu opioid receptor. All compounds displaced the mu opioid receptor binding of [3H]Tyr-D-Ala-Gly-(Me)Phe-Gly-ol in monkey brain membranes ... More
Catalytic properties of lipopolysaccharide (LPS) binding protein. Transfer of LPS to soluble CD14.
AuthorsYu B, Wright SD
JournalJ Biol Chem
PubMed ID8626747
Lipopolysaccharide (LPS) binding protein (LBP) is a lipid transfer protein that catalyzes transfer of LPS monomers from micelles to a binding site on soluble CD14 (sCD14) and transfer of LPS from LPS.sCD14 complexes to HDL particles. To characterize the first of these two reactions, LPS covalently derivatized with the fluorophore, ... More
Fluorescent ligands for studying neuropeptide receptors by confocal microscopy.
AuthorsBeaudet A, Nouel D, Stroh T, Vandenbulcke F, Dal-Farra C, Vincent JP
JournalBraz J Med Biol Res
PubMed ID9921286
This paper reviews the use of confocal microscopy as it pertains to the identification of G-protein coupled receptors and the study of their dynamic properties in cell cultures and in mammalian brain following their tagging with specific fluorescent ligands. Principles that should guide the choice of suitable ligands and fluorophores ... More
Effects of site-directed mutagenesis of basic residues (Arg 94, Lys 95, Lys 99) of lipopolysaccharide (LPS)-binding protein on binding and transfer of LPS and subsequent immune cell activation.
AuthorsLamping N, Hoess A, Yu B, Park TC, Kirschning CJ, Pfeil D, Reuter D, Wright SD, Herrmann F, Schumann RR
JournalJ Immunol
PubMed ID8906845
LPS-binding protein (LBP) is a 60-kDa acute phase glycoprotein capable of binding the LPS of Gram-negative bacteria and facilitating its diffusion. This process is thought to be of potential importance in inflammatory reactions and pathogenic states such as septic shock syndrome. Here, we report on the identification of a LPS ... More
Adsorption of beta-hairpin peptides on the surface of water: a neutron reflection study.
AuthorsLu JR, Perumal S, Powers ET, Kelly JW, Webster JR, Penfold J
JournalJ Am Chem Soc
PubMed ID12656605
Neutron reflectivity has been used to determine the thickness and surface coverage of monolayers of two 14-residue beta-hairpin peptides adsorbed at the air/water interface. The peptides differed only in that one was labeled with a fluorophore, while the other was not. The neutron reflection measurements were mainly made in null ... More
Physical state changes of membrane lipids in human lung adenocarcinoma A(549) cells and their resistance to cisplatin.
AuthorsLiang X, Huang Y
JournalInt J Biochem Cell Biol
PubMed ID12127575
The properties of membrane lipids in sensitive A(549) and resistant A(549)/DDP cells to cis-dichlorodiammine platinum[II] (cisplatin) were examined by combining different approaches. The results showed that fluorescence intensity (deltaF) of Merocyanine 540 (MC540) was 93.5 +/- 21.8 for the sensitive A(549) cells and 49.5 +/- 11.2 for the resistive A(549)/DDP ... More
Structurally related peptide agonist, partial agonist, and antagonist occupy a similar binding pocket within the cholecystokinin receptor. Rapid analysis using fluorescent photoaffinity labeling probes and capillary electrophoresis.
AuthorsDong M, Ding XQ, Pinon DI, Hadac EM, Oda RP, Landers JP, Miller LJ
JournalJ Biol Chem
PubMed ID9988716
The molecular basis of ligand binding to receptors provides important insights for drug development. Here, we explore domains of the cholecystokinin (CCK) receptor that are critical for ligand binding, using a novel series of fluorescent photolabile probes, receptor proteolysis, and rapid high resolution separation of peptide fragments by capillary electrophoresis. ... More
Kinetics of intracellular viral disassembly and processing probed by Bodipy fluorescence dequenching.
AuthorsDa Poian AT, Gomes AM, Coelho-Sampaio T
JournalJ Virol Methods
PubMed ID9506812
A novel method is described for the study of viral disassembly and processing in live cells. Vesicular stomatitis virus (VSV) was labelled with the fluorescent probe Bodipy-FL and the resulting conjugate was 97.6% self-quenched due to fluorescence resonance energy transfer between neighbouring Bodipy molecules. In vitro experiments showed a four-fold ... More
N-Methyl-D-aspartate receptors are clustered and immobilized on dendrites of living cortical neurons.
AuthorsBenke TA, Jones OT, Collingridge GL, Angelides KJ
JournalProc Natl Acad Sci U S A
PubMed ID7689230
The response of nerve cells to synaptic inputs and the propagation of this activation is critically dependent on the cell-surface distribution of ion channels. In the hippocampus, Ca2+ influx through N-methyl-D-aspartate receptors (NMDAR) and/or voltage-dependent calcium channels on dendrites is thought to be critically involved in long-term potentiation, neurite outgrowth, ... More
Metabolism of activated complement component C3 is mediated by the low density lipoprotein receptor-related protein/alpha(2)-macroglobulin receptor.
AuthorsMeilinger M, Gschwentner C, Burger I, Haumer M, Wahrmann M, Szollar L, Nimpf J, Huettinger M
JournalJ Biol Chem
PubMed ID10608878
Complement component 3 (C3) and alpha(2)-macroglobulin evolved from a common, evolutionarily old, ancestor gene. Low density lipoprotein-receptor-related protein/alpha(2)-macroglobulin receptor (LRP/alpha(2)MR), a member of the low density lipoprotein receptor family, is responsible for the clearance of alpha(2)-macroglobulin-protease complexes. In this study, we examined whether C3 has conserved affinity for LRP/alpha(2)MR. Ligand ... More
Targeting of scavenger receptor class B type I by synthetic amphipathic alpha-helical-containing peptides blocks lipopolysaccharide (LPS) uptake and LPS-induced pro-inflammatory cytokine responses in THP-1 monocyte cells.
AuthorsBocharov AV, Baranova IN, Vishnyakova TG, Remaley AT, Csako G, Thomas F, Patterson AP, Eggerman TL
JournalJ Biol Chem
PubMed ID15199068
Human scavenger receptor class B type I, CLA-1, mediates lipopolysaccharide (LPS) binding and internalization (Vishnyakova, T. G., Bocharov, A. V., Baranova, I. N., Chen, Z., Remaley, A. T., Csako, G., Eggerman, T. L., and Patterson, A. P. (2003) J. Biol. Chem. 278, 22771-22780). Because one of the recognition motifs in ... More
Binding and internalization of lipopolysaccharide by Cla-1, a human orthologue of rodent scavenger receptor B1.
AuthorsVishnyakova TG, Bocharov AV, Baranova IN, Chen Z, Remaley AT, Csako G, Eggerman TL, Patterson AP
JournalJ Biol Chem
PubMed ID12651854
Scavenger receptor, class B, type I (SR-BI) mediates selective uptake of high density lipoprotein (HDL) cholesteryl ester. SR-BI recognizes HDL, low density lipoprotein (LDL), exchangeable apolipoproteins, and protein-free lipid vesicles containing negatively charged phospholipids. Lipopolysaccharides (LPS) are highly glycosylated anionic phospholipids contributing to septic shock. Despite significant structural similarities between ... More
Differential internalization of somatostatin in COS-7 cells transfected with SST1 and SST2 receptor subtypes: a confocal microscopic study using novel fluorescent somatostatin derivatives.
AuthorsNouel D, Gaudriault G, Houle M, Reisine T, Vincent JP, Mazella J, Beaudet A
JournalEndocrinology
PubMed ID8977417
A growing body of evidence suggests that neuropeptide binding to G protein-linked receptors may result in internalization of receptor-ligand complexes, followed by intracellular mobilization and degradation of the ligand into its target cells. Because of discrepant results in the literature concerning the occurrence of such a mechanism for the tetradecapeptide ... More
A unique fluorescent phenylalkylamine probe for L-type Ca2+ channels. Coupling of phenylalkylamine receptors to Ca2+ and dihydropyridine binding sites.
AuthorsKnaus HG, Moshammer T, Kang HC, Haugland RP, Glossmann H
JournalJ Biol Chem
PubMed ID1310311
The first fluorescently labeled phenylalkylamine, DMBODIPY-PAA (5-(3-[3-(4,4-difluoro-5,7-dimethyl-3a, 4a-diaza-4-bora-indacen-3-yl)propionamido] phenethyl-N-methylamino)-2-isopropyl-2-(3,4,5-trimethoxyphenyl)-valer onitrile) has been introduced for L-type Ca2+ channel research. DMBODIPY-PAA binds reversibly to L-type Ca2+ channels purified from rabbit skeletal muscle microsomes by wheat germ agglutinin-Sepharose chromatography. In this preparation DMBODIPY-PAA labels 412 pmol of phenylalkylamine receptors/mg of protein with a ... More
Enhancement of leukocyte response to lipopolysaccharide by secretory group IIA phospholipase A2.
AuthorsCai TQ, Thieblemont N, Wong B, Thieringer R, Kennedy BP, Wright SD
JournalJ Leukoc Biol
PubMed ID10380895
Secretory nonpancreatic group IIA phospholipase A2 (sPLA2), a lipolytic enzyme found in plasma, is thought to play an important role in inflammation. In patients with sepsis, a strong positive correlation is observed between the plasma level of sPLA2 and poor clinical outcome in sepsis. We have thus asked whether sPLA2 ... More
Potential role of membrane internalization and vesicle fusion in adhesion of neutrophils in response to lipopolysaccharide and TNF.
AuthorsDetmers PA, Thieblemont N, Vasselon T, Pironkova R, Miller DS, Wright SD
JournalJ Immunol
PubMed ID8955211
Human polymorphonuclear leukocytes (PMN) respond to LPS with strongly increased integrin-mediated adhesion. While the first step of this process has been identified as the interaction of LPS with CD14 on the cell surface, subsequent steps remain to be elucidated. The experiments presented here suggest that monomeric LPS is internalized in ... More
A photocleavable fluorescent nucleotide for DNA sequencing and analysis.
AuthorsLi Z, Bai X, Ruparel H, Kim S, Turro NJ, Ju J
JournalProc Natl Acad Sci U S A
PubMed ID12515853
DNA sequencing by synthesis during a polymerase reaction using laser-induced fluorescence detection is an approach that has a great potential to increase the throughput and data quality of DNA sequencing. We report the design and synthesis of a photocleavable fluorescent nucleoside triphosphate, one of the essential molecules required for the ... More
Four-color DNA sequencing by synthesis on a chip using photocleavable fluorescent nucleotides.
AuthorsSeo TS, Bai X, Kim DH, Meng Q, Shi S, Ruparel H, Li Z, Turro NJ, Ju J
JournalProc Natl Acad Sci U S A
PubMed ID15829588
We report four-color DNA sequencing by synthesis (SBS) on a chip, using four photocleavable fluorescent nucleotide analogues (dGTP-PC-Bodipy-FL-510, dUTP-PC-R6G, dATP-PC-ROX, and dCTP-PC-Bodipy-650) (PC, photocleavable; Bodipy, 4,4-difluoro-4-bora-3alpha,4alpha-diaza-s-indacene; ROX, 6-carboxy-X-rhodamine; R6G, 6-carboxyrhodamine-6G). Each nucleotide analogue consists of a different fluorophore attached to the 5 position of the pyrimidines and the 7 position ... More
Photocleavable fluorescent nucleotides for DNA sequencing on a chip constructed by site-specific coupling chemistry.
AuthorsSeo TS, Bai X, Ruparel H, Li Z, Turro NJ, Ju J
JournalProc Natl Acad Sci U S A
PubMed ID15064393
DNA sequencing by synthesis on a solid surface offers new paradigms to overcome limitations of electrophoresis-based sequencing methods. Here we report DNA sequencing by synthesis using photocleavable (PC) fluorescent nucleotides [dUTP-PC-4,4-difluoro-4-bora-3 alpha,4 alpha-diaza-s-indacene (Bodipy)-FL-510, dCTP-PC-Bodipy-650, and dUTP-PC-6-carboxy-X-rhodamine (ROX)] on a glass chip constructed by 1,3-dipolar azide-alkyne cycloaddition coupling chemistry. Each ... More