Effects of the regulatory light chain phosphorylation of myosin II on mitosis and cytokinesis of mammalian cells.
Authors Komatsu S; Yano T; Shibata M; Tuft R A; Ikebe M;
JournalJ Biol Chem
PubMed ID10944522
'Myosin plays an important role in mitosis, especially during cytokinesis. Although it has been assumed that phosphorylation of regulatory light chain of myosin (RLC) controls motility of mammalian non-muscle cells, the functional significance of RLC phosphorylation remains uninvestigated. To address this problem, we have produced unphosphorylatable RLC (T18A/S19A RLC) and ... More
Spontaneous and ligand-induced trafficking of CXC-chemokine receptor 4.
AuthorsTarasova NI, Stauber RH, Michejda CJ
JournalJ Biol Chem
PubMed ID9632631
'A chimeric protein consisting of CXC-chemokine receptor 4 (CXCR4) and the green fluorescent protein (GFP) was used for studying receptor localization and trafficking in real time in stably transduced HeLa, U-937, CEM, and NIH/3T3 cells. CXCR4-GFP was fully active as a co-receptor in mediating human immunodeficiency virus (HIV) entry. Both ... More
Flow cytometric measurement of fluorescence resonance energy transfer on cell surfaces. Quantitative evaluation of the transfer efficiency on a cell-by-cell basis.
'A method has been developed for the determination of the efficiency (E) of the fluorescence resonance energy transfer between moieties on cell surfaces by use of a computer-controlled flow cytometer capable of dual wavelength excitation. The absolute value of E may be calculated on a single-cell basis. The analysis requires ... More
Proximity of lectin receptors on the cell surface measured by fluorescence energy transfer in a flow system.
AuthorsChan SS, Arndt-Jovin DJ, Jovin TM
JournalJ Histochem Cytochem
PubMed ID374620
'Molecules of the lectin concanavalin A have been labeled separately with the fluorescein and rhodamine chromophores and jointly bound to the surface of transformed Friend erythroleukemia cells. The two dyes constitute an ideal donor-acceptor pair for fluorescence resonance energy transfer thereby permitting the determination of the proximity relationships between bound ... More
Immobilization of concanavalin A receptors during differentiation of neuroblastoma cells.
AuthorsFishman MC, Dragsten PR, Spector I
JournalNature
PubMed ID6261153
'Neuroblastoma cells serve as a useful model of neuronal development because compounds such as dimethyl sulphoxide (DMSO) and dibutyryl cyclic AMP cause them to undergo a process of controlled differentiation in tissue culture, during which they can extend long processes, develop characteristic excitability mechanisms, synthesize neurotransmitters and form synapses. We ... More
Lectin receptor proximity on HL-60 leukemia cells determined by fluorescence energy transfer using flow cytometry.
AuthorsJenis DM, Stepanowski AL, Blair OC, Burger DE, Sartorelli AC
JournalJ Cell Physiol
PubMed ID6594341
'Fluorescence energy transfer using flow cytometric measurements was utilized to determine the proximity of concanavalin A receptors on the surface of HL-60 promyelocytic leukemia cells before and after induction of differentiation. The HL-60 cells were induced to differentiate into granulocytes using dimethylsulfoxide and into macrophages using 12-O-tetradecanoylphorbol-13-acetate. Concanavalin A was ... More
A quantitative analysis of cell allocation to trophectoderm and inner cell mass in the mouse blastocyst.
AuthorsFleming TP
JournalDev Biol
PubMed ID3803716
'The allocation of cells to the trophectoderm and inner cell mass (ICM) in the mouse blastocyst has been examined by labelling early morulae (16-cell stage) with the short-term cell lineage marker yellow-green fluorescent latex (FL) microparticles. FL is endocytosed exclusively into the outside polar cell population and remains autonomous to ... More
Microscopical examination of the localisation patterns of two novel rhodamine derivatives in normal and neoplastic colonic mucosa.
AuthorsAtlamazoglou V, Yova D, Kavantzas N, Loukas S
JournalLasers Med Sci
PubMed ID11702630
'Tissue characterisation by fluorescence imaging, using exogenous fluorophores, is a promising method for cancer detection. Histochemical alterations in the composition of mucins, when neoplastic transformations occur, could be exploited to derive more selective fluoroprobes indicative of early malignant transformation. The aim of this work was to develop and examine tumour ... More
Contractile basis of ameboid movement. VII. The distribution of fluorescently labeled actin in living amebas.
AuthorsTaylor DL, Wang YL, Heiple JM
JournalJ Cell Biol
PubMed ID6893200
'The technique of molecular cytochemistry has been used to follow the distribution of fluorescently labeled actin in living Chaos carolinensis and Amoeba proteus during ameboid movement and various cellular processes. The distribution of 5-iodoacetamidofluorescein-labeled actin was compared with that of Lissamine rhodamine B sulfonyl chloride-labeled ovalbumin microinjected into the same ... More
Lymphocyte mechanical response triggered by cross-linking surface receptors.
AuthorsPasternak C, Elson EL
JournalJ Cell Biol
PubMed ID4038710
'Using a recently developed method (Petersen, N. O., W. B. McConnaughey, and E. L. Elson, 1982, Proc. Natl. Acad. Sci. USA., 79:5327-5331), we have measured changes in the deformability of lymphocytes triggered by cross-linking cell surface proteins. Our study was motivated by two previously demonstrated phenomena: the redistribution ("capping") of ... More
Axonal transport of lectins in the peripheral nervous system.
AuthorsBorges LF, Sidman RL
JournalJ Neurosci
PubMed ID6176698
'The binding and axonal transport of six lectins were studied in the peripheral nervous system of adult mice by an immunocytochemical method. After injection into muscle and subcutaneous sites, lectins known to bind preferentially N-acetylglucosamine or mannose sugars were transported axonally to ventral horn and dorsal root ganglion neurons. Twelve ... More
Application of fluorescently labelled lectins for the visualization and biochemical characterization of polysaccharides in biofilms of Pseudomonas aeruginosa.
AuthorsStrathmann M, Wingender J, Flemming HC
JournalJ Microbiol Methods
PubMed ID12031574
'Fluorescently labelled lectins were used in combination with epifluorescence microscopy and confocal laser scanning microscopy to allow the visualization and characterization of carbohydrate-containing extracellular polymeric substances (EPS) in biofilms of Pseudomonas aeruginosa. A mucoid strain characterized by an overproduction of the exopolysaccharide alginate, and an isogenic, non-mucoid strain were used. ... More
Role for sperm in spatial patterning of the early mouse embryo.
AuthorsPiotrowska K, Zernicka-Goetz M
JournalNature
PubMed ID11206548
'Despite an apparent lack of determinants that specify cell fate, spatial patterning of the mouse embryo is evident early in development. The axis of the post-implantation egg cylinder can be traced back to organization of the pre-implantation blastocyst. This in turn reflects the organization of the cleavage-stage embryo and the ... More
Distribution and mobility of murine histocompatibility H-2Kk antigen in the cytoplasmic membrane.
'The topographical distributions and mobilities of the murine histocompatibility antigen H-2Kk and of concanavalin A (Con A) binding sites have been studied on a murine lymphoma cell line. The spatial distribution of H-2Kk antigens, the average distance between H-2Kk antigens and Con A binding sites, and the separation of different ... More
Spatial and temporal control of nonmuscle myosin localization: identification of a domain that is necessary for myosin filament disassembly in vivo.
AuthorsEgelhoff TT, Brown SS, Spudich JA
JournalJ Cell Biol
PubMed ID1899668
'Myosin null mutants of Dictyostelium are defective for cytokinesis, multicellular development, and capping of surface proteins. We have used these cells as transformation recipients for an altered myosin heavy chain gene that encodes a protein bearing a carboxy-terminal 34-kD truncation. This truncation eliminates threonine phosphorylation sites previously shown to control ... More
Resonance energy transfer microscopy.
AuthorsHerman B
JournalMethods Cell Biol
PubMed ID2648111
Membrane receptor mobility changes by Sendai virus.
AuthorsPringle JR, Preston RA, Adams AE, Stearns T, Drubin DG, Haarer BK, Jones EW
JournalMethods Cell Biol
PubMed ID2476649
Conjugation of lectins with fluorochromes: an approach to histochemical double labeling of carbohydrate components.
AuthorsRoth J, Binder M, Gerhard UJ
JournalHistochemistry
PubMed ID80399
Methodical investigations on the coupling of lectins (Con A, LcL, WGA, RcA) to tetramethylrhodamine isothiocyan ate (TRITC) are reported. 20-microgram of TRITC per mg of lectin were found to be the optimal amount of TRITC for the conjugation. With this fluorochrome: protein ratio conjugates were produced which resulted in a ... More
Unconstrained lateral diffusion of concanavalin A receptors on bulbous lymphocytes.
AuthorsWu ES, Tank DW, Webb WW
JournalProc Natl Acad Sci U S A
PubMed ID6956904
The lateral diffusion coefficient, D, of concanavalin A receptors and receptor complexes on the surface of lymphocytes and RDM4 lymphomas is enhanced by several orders of magnitude to D greater than 5 X 10(-9)cm2/sec by induction of swelling of the cells to bulbous form. Treatments with concanavalin A or 7-nitrobenz-2-oxa-1,3-diazole-phallacidin ... More
Activation of human neutrophil NADPH oxidase and lateral mobility of membrane proteins. A study with crosslinkers.
AuthorsAviram I, Henis YI
JournalBiochim Biophys Acta
PubMed ID6091778
Fluorescence photobleaching recovery was employed to investigate the relationship between the activation of neutrophil NADPH oxidase and lateral mobility of membrane proteins. Treatment of neutrophils with the crosslinking reagent disuccinimidyl suberate (DSS) blocked activation of the respiratory burst without affecting the lateral motion of concanavalin A receptors. Neutrophils treated with ... More
Fluorescence resonance energy transfers measurements on cell surfaces via fluorescence polarization.
AuthorsCohen-Kashi M, Moshkov S, Zurgil N, Deutsch M
JournalBiophys J
PubMed ID12202365
A method has been developed for the determination of the efficiency of fluorescence resonance energy transfer efficiency between moieties located on cell surfaces by performing individual cell fluorescence polarization (FP) measurements. The absolute value of energy transfer efficiency (E) is calculated on an individual cell basis. The examination of this ... More
Schistosoma mansoni: visualization with fluorescent lectins of secretions and surface carbohydrates of living cercariae.
AuthorsLinder E
JournalExp Parasitol
PubMed ID3996523
Attachment of Schistosoma mansoni cercariae was studied during their explorative movements along a glass surface using labeled lectins as markers. Fluorochrome-labeled lectins selectively labeled surface material produced at the cercarial attachment sites and part of the cercarial surface. The deposited secretions reacted with most of the lectins used but differences ... More
Fluorescence photobleaching recovery measurements of surface lateral mobilities on normal and SV40-transformed mouse fibroblasts.
AuthorsEldridge CA, Elson EL, Webb WW
JournalBiochemistry
PubMed ID6246920
Lateral mobilities of fluorescent cell surface probes have been measured on normal (3T3) and transformed (SV3T3) cultured mouse fibroblasts. There is little discernible difference in the mobilities of a lipid analogue (diI), a fluorescent ganglioside derivative (GM1), and tetramethylrhodamine-labeled succinylated concanavalin A. The two cell lines showed expected differences in ... More
Retention of mutant low density lipoprotein receptor in ER leads to ER stress.
Familial hypercholesterolemia (FH) is an autosomal dominant disease caused by mutations in the gene encoding the low density lipoprotein receptor (LDLR). More than 50% of these mutations lead to receptor proteins that are completely or partly retained in the endoplasmic reticulum (ER). The mechanisms involved in the intracellular processing and ... More
Long-range electrostatic trapping of single-protein molecules at a liquid-solid interface.
AuthorsXu XH, Yeung ES
JournalScience
PubMed ID9733506
The motion of single, dye-labeled protein molecules was monitored at various pH and ionic strengths within the 180-nanometer-thick evanescent-field layer at a fused-silica surface. Below the isoelectric point, molecules partitioning into the excitation region increased in number but maintained a random spatial distribution, implying that surface charge can influence the ... More
Peptide-based targeting of fluorophores to organelles in living cells.
AuthorsPap EH, Dansen TB, van Summeren R, Wirtz KW
JournalExp Cell Res
PubMed ID11302694
Peptides carrying organelle-specific import or retention sequences can target the fluorophore BODIPY(581/591) to the nucleus, peroxisomes, endoplasmic reticulum (ER), or the trans-Golgi network (TGN). The peroxisomal peptide contains the PTS1 sequence AKL. For targeting to the ER or TGN, the peptides carry the retention sequences KDEL and SDYQRL, respectively. A ... More
Triplet-state detection of labeled proteins using fluorescence recovery spectroscopy.
AuthorsCorin AF, Blatt E, Jovin TM
JournalBiochemistry
PubMed ID2441744
The experimental procedures for detecting the triplet states of chromophores in solutions (cuvettes) by fluorescence recovery spectroscopy (FRS) are described in detail, together with applications in studies of protein structure and protein-cell interactions in the microsecond to millisecond time domain. The experimental configuration has been characterized by measuring the emission ... More
Rearrangement of Con A and WGA receptors on murine mastocytoma cell membranes: relationship to cell cycle.
AuthorsLustig S, Pluznik DH
JournalJ Cell Physiol
PubMed ID6403559
Incubation of interphase murine mastocytoma cells with fluorescein-labeled wheat germ agglutinin (FITC-WGA) or with rhodamine-labeled concanavalin A (Rh-Con A) shows binding of the lectins to all the cells in a ring form distribution. Treatment of the cells with trypsin before addition of FITC-WGA, or with colchicine before addition of Rh-Con ... More
Fluorescence energy transfer measurements on cell surfaces: a critical comparison of steady-state fluorimetric and flow cytometric methods.
The energy transfer efficiency E was measured between fluorescein-conjugated concanavalin A (Con A) and rhodamine-conjugated Con A bound to homogeneous tissue culture cells, the HK22 murine lymphoma cell line. Results from a flow cytometric energy transfer method (FCET) and two different steady-state fluorimeter methods were compared. The data were found ... More
Fluorescence photobleaching recovery measurements reveal differences in envelopment of Sindbis and vesicular stomatitis viruses.
AuthorsJohnson DC, Schlesinger MJ, Elson EL
JournalCell
PubMed ID6258803
Fluorescence photobleaching recovery (FPR) measurements of virus glycoproteins on the surfaces of cells infected with vesicular stomatitis virus (VSV) and Sindbis virus showed that the VSV glycoprotein (G) remained mobile throughout the infectious cycle, whereas Sindbis virus glycoproteins (E1, E2) were partially mobile early after infection and immobile at later ... More
Extracellular Toll-like receptor 2 region containing Ser40-Ile64 but not Cys30-Ser39 is critical for the recognition of Staphylococcus aureus peptidoglycan.
AuthorsMitsuzawa H, Wada I, Sano H, Iwaki D, Murakami S, Himi T, Matsushima N, Kuroki Y
JournalJ Biol Chem
PubMed ID11522781
Toll-like receptor 2 (TLR2) and CD14 function as pattern recognition receptors for bacterial peptidoglycan (PGN). TLRs and CD14 possess repeats of the leucine-rich motif. To address the role of the extracellular domain of TLR2 in PGN signaling, we constructed CD14/TLR2 chimeras, in which residues 1-356 or 1-323 of CD14 were ... More
Imaging the spatial distribution of membrane receptors during neutrophil phagocytosis.
AuthorsKindzelskii AL, Xue W, Todd RF, Petty HR
JournalJ Struct Biol
PubMed ID7734243
Optical microscopy and image processing have been employed to study the distribution of several cell surface receptors on living human neutrophils during opsonin-dependent and opsonin-independent phagocytosis. Receptors were labeled using fluorescein-, rhodamine-, or AMCA-conjugated F(ab')2 fragments of anti-Fc gamma RIIIB (CD16), anti-CR3 (CD11b/CD18), and anti-uPAR (urokinase-type plasminogen activator receptor) antibodies, ... More
Fluorescence energy transfer-sensitized photobleaching of a fluorescent label as a tool to study donor-acceptor distance distributions and dynamics in protein assemblies: studies of a complex of biotinylated IgM with streptavidin and aggregates of concanavalin A.
AuthorsMekler VM, Averbakh AZ, Sudarikov AB, Kharitonova OV
JournalJ Photochem Photobiol B
PubMed ID9372617
A photokinetic method of detection of fluorescence resonance energy transfer (FRET) between special fluorescent labels is applied to study time-averaged spatial distribution of labeled proteins in protein assemblies. Prolonged irradiation of a sample at the absorption maximum of the energy donor label initiates FRET-sensitized fluorescence photobleaching of the energy acceptor ... More
A fluorescence-based glucose biosensor using concanavalin A and dextran encapsulated in a poly(ethylene glycol) hydrogel.
A fluorescence biosensor is described that is based on a photopolymerized poly(ethylene glycol) (PEG) hydrogel incorporating fluorescein isothiocyanate dextran (FITC-dextran) and tetramethylrhodamine isothiocyanate concanavalin A (TRITC-Con A) chemically conjugated into the hydrogel network using an alpha-acryloyl, omega-N-hydroxysuccinimidyl ester of PEG-propionic acid. In the absence of glucose, TRITC-Con A binds with ... More
Calreticulin affects focal contact-dependent but not close contact-dependent cell-substratum adhesion.
AuthorsFadel MP, Dziak E, Lo CM, Ferrier J, Mesaeli N, Michalak M, Opas M
JournalJ Biol Chem
PubMed ID10329714
We used two cell lines expressing fast (RPEfast) and slow (RPEslow) attachment kinetics to investigate mechanisms of cell-substratum adhesion. We show that the abundance of a cytoskeletal protein, vinculin, is dramatically decreased in RPEfast cells. This coincides with the diminished expression level of an endoplasmic reticulum chaperone, calreticulin. Both protein ... More
Changes in lectin receptor lateral mobilities accompany lymphocyte stimulation.
Changes in lateral mobilities of rabbit lymphocyte membrane components in response to succinyl concanavalin A (S Con A) have been studied by fluorescence photobleaching recovery (FPR). During hrs 0 to 3 after exposure to S Con A, lectin receptor mobilities on both T and B cells fall about 2-fold. Reduced ... More
Flow cytometric measurements of fluorescence energy transfer using single laser excitation.
AuthorsSzöllösi J, Mátyus L, Trón L, Balázs M, Ember I, Fulwyler MJ, Damjanovich S
JournalCytometry
PubMed ID3582060
Flow cytometric energy transfer (FCET) measurements between labeled specific sites of cell surface elements (Szöllosi et al., Cytometry, 5:210-216, 1984) have been extended in a simplified form using a flow cytometer equipped with single excitation beam. This versatile and easily applicable method has several advantages over any nonflow cytometric (i.e., ... More
Immunolocalization of the cellular src protein in interphase and mitotic NIH c-src overexpresser cells.
AuthorsDavid-Pfeuty T, Nouvian-Dooghe Y
JournalJ Cell Biol
PubMed ID1702788
The mouse mAb, mAb 327, that recognizes specifically both pp60v-src and pp60c-src in a wide variety of cells, has been used to determine precisely the various locations of pp60c-src in NIH c-src overexpresser cells, using the technique of immunofluorescence microscopy. In interphase cells, the protein exhibits two main distributions: one ... More
Insulation of a G protein-coupled receptor on the plasmalemmal surface of the pancreatic acinar cell.
Receptor desensitization is a key process for the protection of the cell from continuous or repeated exposure to high concentrations of an agonist. Well-established mechanisms for desensitization of guanine nucleotide-binding protein (G protein)-coupled receptors include phosphorylation, sequestration/internalization, and down-regulation. In this work, we have examined some mechanisms for desensitization of ... More
Visualization of the calcitonin receptor-like receptor and its receptor activity-modifying proteins during internalization and recycling.
AuthorsKuwasako K, Shimekake Y, Masuda M, Nakahara K, Yoshida T, Kitaura M, Kitamura K, Eto T, Sakata T
JournalJ Biol Chem
PubMed ID10882736
Expression of the calcitonin receptor-like receptor (CRLR) and its receptor activity modifying proteins (RAMPs) can produce calcitonin gene-related peptide (CGRP) receptors (CRLR/RAMP1) and adrenomedullin (AM) receptors (CRLR/RAMP2 or -3). A chimera of the CRLR and green fluorescent protein (CRLR-GFP) was used to study receptor localization and trafficking in stably transduced ... More
Concanavalin A-induced redistribution of surface receptors in Acanthamoeba castellanii at different growth phases.
AuthorsPaatero GI, Wikström L, Isomaa B
JournalEur J Cell Biol
PubMed ID3229417
Concanavalin A (ConA)-induced redistribution of surface receptors has been studied in Acanthamoeba castellanii at different growth phases utilizing double fluorescent techniques and transmission electron microscopy. When the amoebae were incubated with 2 micrograms and 10 micrograms tetramethylrhodamine isothiocyanate (TRITC)-ConA/ml for 4 min and 15 min at 28 degrees C the ... More
Functions of microtubules in the Saccharomyces cerevisiae cell cycle.
AuthorsJacobs CW, Adams AE, Szaniszlo PJ, Pringle JR
JournalJ Cell Biol
PubMed ID3049620
We used the inhibitor nocodazole in conjunction with immunofluorescence and electron microscopy to investigate microtubule function in the yeast cell cycle. Under appropriate conditions, this drug produced a rapid and essentially complete disassembly of cytoplasmic and intranuclear microtubules, accompanied by a rapid and essentially complete block of cellular and nuclear ... More
Determining the extent of labeling for tetramethylrhodamine protein conjugates.
AuthorsMeadows DL, Shafer JS, Schultz JS
JournalJ Immunol Methods
PubMed ID1719100
A new, relatively simple, spectrophotometric technique has been developed which is useful for accurately determining the extent of chromophore labeling of proteins. Often the absorbance spectra and extinction coefficients of dye/protein conjugates are strongly affected by changes in the chromophore microenvironment that may occur at high dye/protein ratios. In the ... More
Rab7: a key to lysosome biogenesis.
AuthorsBucci C, Thomsen P, Nicoziani P, McCarthy J, van Deurs B
JournalMol Biol Cell
PubMed ID10679007
The molecular machinery behind lysosome biogenesis and the maintenance of the perinuclear aggregate of late endocytic structures is not well understood. A likely candidate for being part of this machinery is the small GTPase Rab7, but it is unclear whether this protein is associated with lysosomes or plays any role ... More
The NK1 receptor localizes to the plasma membrane microdomains, and its activation is dependent on lipid raft integrity.
AuthorsMonastyrskaya K, Hostettler A, Buergi S, Draeger A
JournalJ Biol Chem
PubMed ID15590676
The spatial targeting of receptors to discrete domains within the plasma membrane allows their preferential coupling to specific effectors, which is essential for rapid and accurate discrimination of signals. Efficiency of signaling is further increased by protein and lipid segregation within the plasma membrane. We have previously demonstrated the importance ... More
Microrheology reveals microscale viscosity gradients in planktonic systems.
Authors
JournalProc Natl Acad Sci U S A
PubMed ID33443173
Structural changes of Chlamydomonas reinhardtii cells during lipid enrichment and after solvent exposure.
Authors
JournalData Brief
PubMed ID29845099
Resilience of bacterial quorum sensing against fluid flow.
Authors
JournalSci Rep
PubMed ID27650454
Impact of pulsed electric fields and mechanical compressions on the permeability and structure of Chlamydomonas reinhardtii cells.
Authors
JournalSci Rep
PubMed ID32060324
Fatty acylation enhances the cellular internalization and cytosolic distribution of a cystine-knot peptide.
Authors
JournaliScience
PubMed ID34712919
Dual-species biofilms of Streptococcus mutans and Candida albicans exhibit more biomass and are mutually beneficial compared with single-species biofilms.
Authors
JournalJ Oral Microbiol
PubMed ID31681463
C1q/TNF-Related Protein 3 Prevents Diabetic Retinopathy via AMPK-Dependent Stabilization of Blood-Retinal Barrier Tight Junctions.
Authors
JournalCells
PubMed ID35269401
Adaptive selection drives TRPP3 loss-of-function in an Ethiopian population.
Authors
JournalSci Rep
PubMed ID33268808
Quantitation of yeast cell-cell fusion using multicolor flow cytometry.
Authors
JournalCytometry A
PubMed ID26033928
Social motility of biofilm-like microcolonies in a gliding bacterium.