Autoantigens targeted in systemic lupus erythematosus are clustered in two populations of surface structures on apoptotic keratinocytes.
AuthorsCasciola-Rosen LA, Anhalt G, Rosen A
JournalJ Exp Med
PubMed ID7511686
'Systemic lupus erythematosus is a multisystem autoimmune disease in which the autoantibody response targets a variety of autoantigens of diverse subcellular location. We show here that these autoantigens are clustered in two distinct populations of blebs at the surface of apoptotic cells. The population of smaller blebs contains fragmented endoplasmic ... More
The G protein of vesicular stomatitis virus has free access into and egress from the smooth endoplasmic reticulum of UT-1 cells.
AuthorsBergmann JE, Fusco PJ
JournalJ Cell Biol
PubMed ID2155242
'We have investigated the role of the smooth endoplasmic reticulum (SER) of UT-1 cells in the biogenesis of the glycoprotein (G) of vesicular stomatitis virus (VSV). Using immunofluorescence microscopy, we observed the wild type G protein in the SER of infected cells. When these cells were infected with the mutant ... More
Spectroscopic assays for measuring quantities of erythrocyte membrane "halves".
AuthorsFisher KA
JournalJ Cell Biol
PubMed ID7056801
'The quantities of outer and inner"halves" produced by freeze-fracturing human erythrocyte membranes have been measured by visible and fluorescence spectroscopy. Assays have been developed that are based on the use of two membrane surface markers: hemoglobin (Hb), a native marker for the cytoplasmic side of the membrane, and fluoresceinated concanavalin ... More
Concanavalin A-binding glycoproteins in the subcommissural and the pineal organ of the sheep (Ovis aries). A fluorescence-microscopic and electrophoretic study.
AuthorsMeiniel R, Molat JL, Meiniel A
JournalCell Tissue Res
PubMed ID3757020
'Glycoproteins rich in mannosyl or glucosyl residues were analyzed in the subcommissural organ (SCO) and the pineal organ of the sheep (Ovis aries). By use of concanavalin A labelled with fluorescein isothiocyanate, fluorescent material was found both in ependymal and hypendymal cells of the SCO. In the pineal organ, either ... More
The role of lateral migration in the formation of acetylcholine receptor clusters induced by basic polypeptide-coated latex beads.
AuthorsPeng HB, Zhao DY, Xie MZ, Shen ZW, Jacobson K
JournalDev Biol
PubMed ID2909404
'During the formation of the neuromuscular junction, the nerve induces the clustering of acetylcholine receptors (AChR) in the postsynaptic membrane. This process can be mimicked by treating cultured Xenopus myotomal muscle cells with basic polypeptide-coated latex beads. Using this bead-muscle coculture system, we examined the role of lateral migration of ... More
Scatchard analysis of fluorescent concanavalin A binding to lymphocytes.
AuthorsGordon IL
JournalCytometry
PubMed ID7587709
'Standard Scatchard analysis of ligand binding to cell receptors requires the use of isotopes and is imprecise at low ligand concentrations. To evaluate the feasibility of Scatchard analysis via fluorescence flow cytometry, the binding of fluorescein isothiocyanate-derivatized concanavalin A (FITC-ConA) to murine lymphocytes at 4 degrees C was compared to ... More
Concanavalin A derivatives with altered biological activities.
AuthorsGunther GR, Wang JL, Yahara I, Cunningham BA, Edelman GM
JournalProc Natl Acad Sci U S A
PubMed ID4515602
'Chemical derivatization of tetrameric concanavalin A (Con A) with succinic anhydride or acetic anhydride converts the protein to a dimeric molecule without altering its carbohydrate-binding specificity. At low concentrations, the dose-response curves for the mitogenic stimulation of mouse spleen cells by native Con A and succinyl-Con A are similar. Above ... More
Use of carbohydrate probes in conjunction with fluorescence-activated cell sorting to select mutant cell lines of Chlamydomonas with defects in cell surface glycoproteins.
AuthorsBloodgood RA, Salomonsky NL, Reinhart FD
JournalExp Cell Res
PubMed ID3691676
'Two carbohydrate-binding probes (the lectin concanavalin A and the anti-carbohydrate monoclonal antibody FMG-1) have been utilized in conjunction with fluorescence-activated cell sorting to select cell lines of Chlamydomonas reinhardtii that contain defects in cell surface-exposed glycoproteins. Two very different selection strategies (sorting cells with the lowest binding for the FMG-1 ... More
Acetylcholine receptor aggregation parallels the deposition of a basal lamina proteoglycan during development of the neuromuscular junction.
AuthorsAnderson MJ, Klier FG, Tanguay KE
JournalJ Cell Biol
PubMed ID6386827
'To determine the time course of synaptic differentiation, we made successive observations on identified, nerve-contacted muscle cells developing in culture. The cultures had either been stained with fluorescent alpha-bungarotoxin, or were maintained in the presence of a fluorescent monoclonal antibody. These probes are directed at acetylcholine receptors (AChR) and a ... More
Inhibition of receptor-mediated but not fluid-phase endocytosis in polymorphonuclear leukocytes.
AuthorsDaukas G, Zigmond SH
JournalJ Cell Biol
PubMed ID4055891
'We have found that hypertonic medium inhibited the receptor-mediated uptake of the chemotactic peptide N-formylnorleucylleucylphenylalanine without affecting fluid-phase endocytosis by polymorphonuclear leukocytes (PMNs). Morphological and biochemical evidence demonstrated that cells in hypertonic medium did not accumulate peptide in a receptor-mediated manner. However, the cells continued to form endosomes containing fluid-phase ... More
Concanavalin A induced apoptosis in murine macrophage PU5-1.8 cells through clustering of mitochondria and release of cytochrome c.
AuthorsSuen YK, Fung KP, Choy YM, Lee CY, Chan CW, Kong SK
JournalApoptosis
PubMed ID11227218
'Concanavalin A (ConA), normally a mitogen of T-lymphocytes, was found to be a cell cycle-independent apoptosis-inducing agent in cultured murine macrophage PU5-1.8 cells. This assertion is based on the following observations: (1) ConA increased the number of cells with hypodiploid DNA in a dose dependent manner as revealed by flow ... More
Mobility of concanavalin A receptors and distribution of cytoplasmic actin in odontogenic epithelial and mesenchymal cells.
AuthorsPrime SS, Toh BH
JournalJ Cell Sci
PubMed ID363732
'The distribution of concanavalin A (Con A) surface receptors and cytoplasmic actin in the same cell was studied in monolayer cultures of 2 odontogenic epithelial cells of different developmental age and in ecto-mesenchymal cells derived from the same tooth germ. Con A receptors were demonstrated by fluorescein-isothiocyanate-labelled Con A (FITC-Con ... More
Optical imaging fiber-based single live cell arrays: a high-density cell assay platform.
AuthorsBiran I, Walt DR
JournalAnal Chem
PubMed ID12141663
'A high-density, ordered array containing thousands of microwells is fabricated on an optical imaging fiber. Each individually addressable microwell is used to accommodate a single living cell. A charged coupled device (CCD) detector is employed to monitor and spatially resolve the fluorescence signals obtained from each individual cell, allowing simultaneous ... More
Reduced lateral mobility of a fluorescent lipid analog in cell membranes of rat fibroblasts transformed by simian virus 40.
AuthorsKawasaki Y, Zundel JL, Miyakawa T, Sakaguchi K
JournalFEBS Lett
PubMed ID2540041
'In order to study the difference between normal and transformed cells, lateral motion of fluorescent molecules embedded into cell membranes of rat clonal fibroblasts and its SV40-transformed derivative cells was measured by the FPR technique. The lateral diffusion coefficient of a fluorescent fatty acid analog, F18, was smaller in transformed ... More
A rapid method for quantitating lymphocyte receptor capping: capping defect in AIDS patients.
AuthorsChanh TC, Alderete BE
JournalJ Virol Methods
PubMed ID1702445
'Capping of concanavalin A (Con A) and anti-Leu-8 (L8) receptors on human peripheral blood mononuclear (PBM) membranes was studied utilizing fluorochrome-conjugated ligands and flow cytometric analysis. Histogram profiles of fluorescent intensities consistently revealed a time-dependent decrease in numbers of brightly fluorescing events concurrent with an increase in numbers of dimly ... 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
Immunofluorescent quantification of ribonucleotide reductase M1 subunit and correlation with DNA content by flow cytometry.
AuthorsMann GJ, Dyne M, Musgrove EA
JournalCytometry
PubMed ID3311659
'The cytoplasmic enzyme ribonucleotide reductase is essential for DNA synthesis, and its activity is strongly correlated with cellular proliferation. This paper describes a flow cytometric technique for the simultaneous measurement of DNA content and the M1 subunit of ribonucleotide reductase. Data are presented for cycling cultured human leukemic lymphoblasts in ... More
Vitronectin at sites of cell-substrate contact in cultures of rat myotubes.
AuthorsBaetscher M, Pumplin DW, Bloch RJ
JournalJ Cell Biol
PubMed ID2426279
'Affinity-purified antibodies to the serum glycoprotein, vitronectin, were used to study sites of cell-substrate contact in cultures of rat myotubes and fibroblasts. Cells were removed from the substrate by treatment with saponin, leaving fragments of plasma membrane attached to the glass coverslip. When stained for vitronectin by indirect immunofluorescence, large ... More
Fluorescence-labeled lectins, glycoconjugates, and the development of the mouse AOP.
AuthorsVanden Hoek TL, Goossens W, Knepper PA
JournalInvest Ophthalmol Vis Sci
PubMed ID3557857
'The development of the aqueous outflow pathway (AOP) in early postnatal mouse eyes was examined for the presence of a variety of lectin receptors using fluorescein isothiocyanate (FITC) conjugated lectins, 1 micron araldite plastic sections, and computer-aided fluorescence photography. The trabecular meshwork anlage (days 1-4) was characterized by the presence ... More
Surface carbohydrates of procyclic forms of African trypanosomes studied using fluorescence activated cell sorter analysis and agglutination with lectins.
AuthorsMutharia LM, Pearson TW
JournalMol Biochem Parasitol
PubMed ID3106807
'Living culture form procyclics of Trypanosoma brucei brucei, T.b. rhodesiense, T.b. gambiense, T. congolense and T. simiae were tested for binding of eight different lectins. The binding of fluorescein isothiocyanate (FITC)-conjugated lectins was measured using a fluorescence activated cell sorter (FACS) and by agglutination with unlabelled lectins. Five of the ... 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
Reversion from transformed to normal phenotype by inhibition of protein synthesis in rat kidney cells infected with a temperature-sensitive mutant of Rous sarcoma virus.
AuthorsAsh JF, Vogt PK, Singer SJ
JournalProc Natl Acad Sci U S A
PubMed ID185620
'By the use of a rat kidney cell line infected with a temperature-sensitive Rous sarcoma virus, we have shown that, at permissive temperatures where the cells are transformed, concanavalin A induces a clustering of its cell membrane receptors into patches, and the intracellular smooth muscle myosin-like protein is in a ... More
Fluorochrome-coupled lectins reveal distinct cellular domains in human epidermis.
AuthorsVirtanen I, Kariniemi AL, Holthöfer H, Lehto VP
JournalJ Histochem Cytochem
PubMed ID3512692
'The distribution of saccharide moieties in human interfollicular epidermis was studied with fluorochrome-coupled lectins. In frozen sections Concanavalin A (Con A), Lens culinaris agglutinin (LCA), Ricinus communis agglutinin I (RCAI), and wheat germ agglutinin (WGA) stained intensively both dermis and viable epidermal cell layers, whereas peanut agglutinin (PNA) bound only ... More
Signal- and energy-dependent nuclear transport of SV40 Vp3 by isolated nuclei. Establishment of a filtration assay for nuclear protein import.
AuthorsDean DA, Kasamatsu H
JournalJ Biol Chem
PubMed ID8106464
'Nuclear transport signal (NTS)-containing proteins are transported into the nucleus through the nuclear pore complex by a mechanism that is not well understood. To better characterize the mechanisms of transport, we have established an homologous in vitro system using an NTS-containing structural protein of simian virus 40 (SV40) and isolated ... More
Evaluation of mouse sperm acrosomal status and viability by flow cytometry.
AuthorsTao J, Critser ES, Critser JK
JournalMol Reprod Dev
PubMed ID8257567
'A procedure was developed to evaluate mouse sperm acrosomal status and viability simultaneously utilizing flow cytometry. Four fluorescein isothiocyanate (FITC)-conjugated lectins, peanut agglutinin (PNA), concanavalin agglutinin (ConA), Pisum sativum agglutinin (PSA), and soybean agglutinin (SBA), were investigated, with PNA providing the greatest sensitivity and specificity in distinguishing acrosome-present and acrosome-absent ... 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
LM fibroblast plasma membrane subfractionation by affinity chromatography on con A-sepharose.
AuthorsSchroeder F, Fontaine RN, Kinden DA
JournalBiochim Biophys Acta
PubMed ID7126576
'Affinity chromatography was used to determine the heterogeneity and orientation of plasma membrane vesicles isolated from LM fibroblasts subjected to Dounce homogenization. Two plasma membrane subfractions were obtained by Con A-Sepharose affinity chromatography of LM fibroblast plasma membranes prepared by Dounce homogenization. The desmosterol-phospholipid molar ratio, the phospholipid composition, and ... More
'In this study, we have identified and partially characterized a mouse T-lymphoma ryanodine receptor on a unique type of internal vesicle which bands at the relatively light density of 1.07 g/ml. Analysis of the binding of [3H]ryanodine to these internal vesicles reveals the presence of a single, low affinity binding ... More
Microdomains of distinctive glycoprotein composition in the kidney proximal tubule brush border.
'Two membrane proteins, maltase and gp330 (the pathogenic antigen of Heymann nephritis), present in the proximal tubule brush border have recently been independently purified and found to be large glycoproteins of similar molecular weight (Mr = approximately 300,000) by SDS PAGE. To determine the relationship between the two, monoclonal antibodies ... More
Stimulation of cortical actin polymerization in the sea urchin egg cortex by NH4Cl, procaine and urethane: elevation of cytoplasmic pH is not the common mechanism of action.
AuthorsBegg DA, Wong GK, Hoyle DH, Baltz JM
JournalCell Motil Cytoskeleton
PubMed ID8913642
'Previous studies have demonstrated that the penetrating weak base NH4Cl and the anesthetics procaine and urethane disrupt the normal attachment of cortical granules to the cortex of the sea urchin egg. Hylander and Summers (1981: Dev. Biol. 86:1-11) hypothesized that this effect may be caused by a pH-induced polymerization of ... More
Expression of several adhesive macromolecules (N-CAM, L1, J1, NILE, uvomorulin, laminin, fibronectin, and a heparan sulfate proteoglycan) in embryonic, adult, and denervated adult skeletal muscle.
AuthorsSanes JR, Schachner M, Covault J
JournalJ Cell Biol
PubMed ID3511069
'Levels of the neural cell adhesion molecule N-CAM in muscle are regulated in parallel with the susceptibility of muscle to innervation: N-CAM is abundant on the surface of early embryonic myotubes, declines in level as development proceeds, reappears when adult muscles are denervated or paralyzed, and is lost after reinnervation ... More
Lectin binding to newt epidermis: fluorescent localization and effects on motility.
AuthorsMahan JT, Donaldson DJ
JournalJ Exp Zool
PubMed ID3091758
'The ability of seven lectins to bind to newt epidermal cells and influence their motility was examined. Of the seven fluoresceinated lectins applied to frozen sections containing intact newt skin and migrating epidermis (wound epithelium), only Con A (concanavalin A), WGA (wheat germ agglutinin), and PNA (peanut agglutinin) produced detectable ... More
Lectin histochemistry of classic and spermatocytic seminoma.
AuthorsLee MC, Talerman A, Oosterhuis JW, Damjanov I
JournalArch Pathol Lab Med
PubMed ID3929746
'Six spermatocytic and 13 classic seminomas were studied histochemically with fluorescein isothiocyanate-labeled lectins. Wheat-germ lectin and succinyl concanavalin A reacted with all 19 tumors, whereas other lectins bound to some, but never to all, tumors in either group. Soybean lectin reacted with seven of 13 classic seminomas but with none ... More
Interaction of tumour promoters with epithelial cells in culture. An immunofluorescence study.
AuthorsKellie S, Holme TC, Bissell MJ
JournalExp Cell Res
PubMed ID3930274
'12-O-tetradecanoyl phorbol-13-acetate (TPA) has a profound and rapid influence on the cytoskeleton of Madin-Darby Canine Kidney (MDCK) cells. Within 10 min, TPA induces a rapid change in morphology, from a flat, cuboidal state to a rounded or elongated morphology in which the cell membranes become convoluted. Concomitant with this morphological ... More
Preparation of a high-affinity photolabeling reagent for the Gal/GalNAc lectin of mammalian liver: demonstration of galactose-combining sites on each subunit of rabbit hepatic lectin.
AuthorsLee RT, Lee YC
JournalBiochemistry
PubMed ID2432926
'On the basis of the knowledge that the D-galactose/N-acetyl-D-galactosamine-specific lectin of rabbit liver can tolerate a large group on the C-6 hydroxyl group of a galactoside [Lee, R. T. (1982) Biochemistry 21, 1045-1050], we prepared a high-affinity photolabeling reagent for this lectin from a triantennary glycopeptide fraction of asialofetuin. The ... More
Diminished calcium influx in lectin-stimulated T cells from old mice.
AuthorsMiller RA, Jacobson B, Weil G, Simons ER
JournalJ Cell Physiol
PubMed ID3497930
'T lymphocytes from aged donors function poorly, but the biochemical basis for the defect remains uncertain. We tested the hypothesis that T cells from old mice had a diminished ability to transmit extracellular signals into the cytoplasm, by measuring intracellular free calcium concentrations (Cai) in T cells stimulated by the ... More
Immunofluorescent localization of a 39,000-dalton substrate of tyrosine protein kinases to the cytoplasmic surface of the plasma membrane.
AuthorsNigg EA, Cooper JA, Hunter T
JournalJ Cell Biol
PubMed ID6304112
'The intracellular distribution of p39, a 39,000-dalton substrate for a number of tyrosine protein kinases, has been determined by indirect immunofluorescence microscopy. No binding of anti-p39 antibodies to intact cells was observed, indicating that this protein is not accessible to antibody on the cell surface. Following detergent permeabilization of formaldehyde-fixed ... More
BED1, a gene encoding a galactosyltransferase homologue, is required for polarized growth and efficient bud emergence in Saccharomyces cerevisiae.
AuthorsMondésert G, Reed SI
JournalJ Cell Biol
PubMed ID8567719
'The ellipsoidal shape of the yeast Saccharomyces cerevisiae is the result of successive isotropic/apical growth switches that are regulated in a cell cycle-dependent manner. It is thought that growth polarity is governed by the remodeling of the actin cytoskeleton that is itself under the control of the cell cycle machinery. ... More
Electric field-directed fibroblast locomotion involves cell surface molecular reorganization and is calcium independent.
AuthorsBrown MJ, Loew LM
JournalJ Cell Biol
PubMed ID7929557
'Directional cellular locomotion is thought to involve localized intracellular calcium changes and the lateral transport of cell surface molecules. We have examined the roles of both calcium and cell surface glycoprotein redistribution in the directional migration of two murine fibroblastic cell lines, NIH 3T3 and SV101. These cell types exhibit ... More
Binding of mitogenic plant lectins to human lymphocytes. Flow cytometric analysis.
AuthorsSerke S, Neubauer A, Van Lessen A
JournalJ Immunol Methods
PubMed ID2527272
'Several plant lectins, such as phytohaemagglutinin (PHA), pokeweed mitogen (PWM), concanavalin A (ConA), Maclura pumifera (MPA) and Pisum sativum agglutinin (PSA), are potent mitogens for human lymphocytes. The pattern of activation induced, however, is not uniform for all mitogenic lectins. The different biological effects following lectin activation of human lymphocytes ... More
Isolation of a cDNA encoding human lysophosphatidic acid phosphatase that is involved in the regulation of mitochondrial lipid biosynthesis.
AuthorsHiroyama M, Takenawa T
JournalJ Biol Chem
PubMed ID10506173
'In this study, we isolated cDNA encoding lysophosphatidic acid (LPA) phosphatase (LPAP). The amino acid sequence deduced from the cDNA encoding LPAP had 421 residues including a putative signal peptide and was homologous to acid phosphatase, especially at the active site. Human LPAP had 28.5% amino acid identity to human ... More
Lectin receptors of rods and cones. Visualization by fluorescent label.
AuthorsBridges CD
JournalInvest Ophthalmol Vis Sci
PubMed ID6969714
'The binding of eight fluorescence-labeled lectins to visual cells was investigated. Concanavalin A (Con A) (specific for oligosaccharides containing mannose) bound evenly to the surfaces of rod outer segments (ROSs) from frog, cattle, goldfish, and turtle (Pseudemys). In frog, binding was observed on the inner segment, with intensifications at the ... More
Differential endocytosis of fluorescein iso-thiocyanate-concanavalin A by normal and chronic myeloid leukemic granulocytes.
AuthorsZingde SM, Anklesaria PN, Advani SH, Bhisey AN, Gothoskar BP
JournalBlut
PubMed ID3475138
'Isolated granulocytes from normal individuals and patients suffering from chronic myeloid leukemia (CML) displayed different fluorescent patterns on treatment with fluorescein isothiocyanate concanavalin A (Fl-Con A). The ligand was internalized by 86% of the normal granulocytes, while 80% of the leukemic granulocytes exhibited Fl-Con A localized on the cell periphery. ... More
Adhesion of glycosaminoglycan-deficient chinese hamster ovary cell mutants to fibronectin substrata.
AuthorsLeBaron RG, Esko JD, Woods A, Johansson S, Höök M
JournalJ Cell Biol
PubMed ID3346331
'We have examined the role of cell surface glycosaminoglycans in fibronectin-mediated cell adhesion by analyzing the adhesive properties of Chinese hamster ovary cell mutants deficient in glycosaminoglycans. The results of our study suggest that the absence of glycosaminoglycans does not affect the initial attachment and subsequent spreading of these cells ... More
Membrane changes in neural target cells studied with fluorescent lectin probes.
AuthorsGualandris L, Rougé P, Duprat AM
JournalJ Embryol Exp Morphol
PubMed ID6655431
'The competent ectoderm of Pleurodeles waltl comprises two cell layers with characteristic differences in their morphology, their composition and the molecular arrangement of the various constituents. The use of labelled lectin probes for observations of ectoderm tissue in vitro with u.v. microscopy (epi-illumination) and the quantification of the results show ... More
Studies on factor VIII-related protein. III. Size distribution and carbohydrate content of human and bovine factor VIII.
AuthorsFurlan M, Perret BA, Beck EA
JournalBiochim Biophys Acta
PubMed ID534647
'Human and bovine factor VIII were isolated from cryoprecipitate of fresh frozen plasma by gel filtration on Sepharose CL-2B. The elution diagrams and SDS-agarose electrophoretic analysis of eluted fractions show no significant differences in the size-distribution of factor VIII aggregates between the two species. Agarose gels were stained for carbohydrate ... More
Preparation of fluorescent derivatives of lipases and their use in fluorescence energy transfer studies in hydrocarbon/water interfaces.
AuthorsRoberts GA, Tombs MP
JournalBiochim Biophys Acta
PubMed ID3113482
'Fluorescein isothiocyanate reacted with a chromobacter and pseudomonad lipase to yield mono-substituted, fully active, enzymes. With the carbocyanine dye 1,1''-dioctadecyl-3,3,3'',3''-tetramethylindocarbocyanine perchlorate (DiI) in the non-aqueous phase, fluorescence energy transfer was used to follow the lipase and similarly labelled model proteins in and out of the interface in heptane, and heptane/di-O-palmitoyl-rac-glycerol ... More
Photobleaching recovery studies of membrane events accompanying lectin stimulation of rabbit lymphocytes.
AuthorsLeuther MD, Barisas BG, Peacock JS, Krakauer H
JournalBiochem Biophys Res Commun
PubMed ID314290
A new class of model glycolipids: synthesis, characterization, and interaction with lectins.
Morphogenesis in the yeast cell cycle: regulation by Cdc28 and cyclins.
AuthorsLew DJ, Reed SI
JournalJ Cell Biol
PubMed ID8449978
'Analysis of cell cycle regulation in the budding yeast Saccharomyces cerevisiae has shown that a central regulatory protein kinase, Cdc28, undergoes changes in activity through the cell cycle by associating with distinct groups of cyclins that accumulate at different times. The various cyclin/Cdc28 complexes control different aspects of cell cycle ... 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
Rapid visualization of three common fungi using fluorescein-conjugated lectins.
AuthorsRobin JB, Arffa RC, Avni I, Rao NA
JournalInvest Ophthalmol Vis Sci
PubMed ID3514531
'The feasibility of using fluorescein-conjugated lectins to visualize and differentiate three fungi commonly involved in ophthalmic mycoses was evaluated. Using a panel of fluorescein-conjugated lectins, Candida albicans, Aspergillus fumigatus, and Fusarium solani were rapidly and reproducibly visualized in in vitro culture isolates, as well as in tissue samples and fixed ... More
Fluorescent antibodies and lectins stain intracellular structures in fixed cells treated with nonionic detergent.
AuthorsLaurila P, Virtanen I, Wartiovaara J, Stenman S
JournalJ Histochem Cytochem
PubMed ID207770
'Nonionic detergent (NP40) treatment of paraformaldehyde-fixed normal and SV40-transformed human fibroblasts resulted in intracellular penetration of two chosen fluorescent antibodies and Concanavalin A (Con A). After the detergent treatment nuclear SV40 T antigen, cytoplasmic fibronectin glycoprotein and Con A binding sites could be visualized in fluorescence microscopy. The lowest NP40 ... More
Protein blotting: detection of proteins with colloidal gold, and of glycoproteins and lectins with biotin-conjugated and enzyme probes.
AuthorsRohringer R, Holden DW
JournalAnal Biochem
PubMed ID2984955
'Methods to detect "native" proteins immobilized on nitrocellulose membranes in spot tests or on blots prepared from polyacrylamide slab gels after electrophoretic separation are described. Gold sols were found to be useful as general stains for proteins: They are polychromatic, yield an indelible record, and are complementary to india ink ... More
Wet cleaving of cells: a method to introduce macromolecules into the cytoplasm. Application for immunolocalization of cytosol-exposed antigens.
AuthorsBrands R, Feltkamp CA
JournalExp Cell Res
PubMed ID3132398
'A new method, called "wet cleaving," has been introduced to allow direct exposure of cytoplasm to externally supplied macromolecules by mechanical rupture of the plasma membrane. Monolayers of adherent cells or poly-L-lysine-attached suspension cells are overlayed with nitrocellulose sheets. By subsequent removal of the sheets, cells are cleaved, thereby exposing ... More
Use of lectins and polyethylene glycol for fusion of glycolipid-containing liposomes with eukaryotic cells.
AuthorsSzoka F, Magnusson KE, Wojcieszyn J, Hou Y, Derzko Z, Jacobson K
JournalProc Natl Acad Sci U S A
PubMed ID6940182
'Efficient fusion of phospholipid vesicles with monolayer cultures of eukaryotic cells was accomplished by attaching glycolipid-containing vesicles to the cell surface by using a lectin displaying binding for both the cell surface and the glycolipid, followed by treatment with polyethylene glycol. Fusion was inferred from the transfer of fluorescent lipid ... More
Flow cytometric analyses of lectin binding to rat alveolar macrophages.
AuthorsSaunders GC, Steinkamp JA, Lehnert BE
JournalCytometry
PubMed ID3123174
'The binding characteristics of ten FITC-labeled plant lectins (Con-A, MPA, BPA, PNA, WGA, SBA, UEA-I, DBA, GS-I, GS-II) to lavaged rat alveolar macrophages were assessed by flow cytometry. The alveolar macrophages (AM) were incubated with varying concentrations of each lectin in a pinocytosis-inhibiting buffer. In addition to measuring lectin-associated green ... More
Comparative responses of normal and malignant mouse mammary cells to modulation of surface properties.
'We previously demonstrated that epithelial cells growing in primary cultures derived from normal and neoplastic mouse mammary tissues are indistinguishable on the basis of morphology, surface topography, agglutinability by concanavalin A, or of number, cytoplasmic location, or organization of microtubules and actin-containing microfilaments. In the present study, the modulation and ... More
Spectroscopic and microscopic studies of buffalo-bull (Bubalus bubalis) spermatozoa.
AuthorsBawa SR, Govil G, Dhingra MS, Werner G, Bains HK
JournalJ Submicrosc Cytol Pathol
PubMed ID8384930
'We have examined the epididymal (caput, corpus and cauda) and ejaculated spermatozoa of bufallo-bull (Bubalus bubalis) employing microscopic and spectroscopic techniques. Fluorescein isothiocyanate conjugated lectins namely concanavalin A (Con A), Dolichos biflorus (DBA), Maclura pomifera (MPA), peanut agglutinin (PNA), soybean agglutinin (SBA) and wheat germ agglutinin (WGA) were used to ... More
Lectins and heparin-binding features of human spermatozoa as analyzed by flow cytometry.
AuthorsVasquez JM, Magargee SF, Kunze E, Hammerstedt RH
JournalAm J Obstet Gynecol
PubMed ID2124086
'Sperm cell surface characteristics may serve as a useful diagnostic feature for the evaluation of seminal samples, but to date no useful analytical method(s) is available for their routine quantitative analysis. Fluorescein isothiocyanate-conjugated lectins, proteins with unique and specific saccharide-binding characteristics, and heparin are potentially useful markers for such an ... 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
The intracellular movement of endocytic vesicles in cultured granulosa cells.
AuthorsHerman B, Albertini DF
JournalCell Motil
PubMed ID6132682
'Ligand binding to cell surface receptors induces rapid internalization of ligand-receptor complexes by receptor mediated endocytosis. We have examined the intracellular movement of endocytic vesicles, induced by the lectin concanavalin A (Con A), in cultured rat ovarian granulosa cells using fluorescence and electron microscopy. Within 20 minutes of ligand treatment ... More
Use of fluorescent probes to monitor propranolol effects on the murine splenic lymphocyte.
AuthorsAudus KL, Johnson DL, Gordon MA
JournalJ Immunopharmacol
PubMed ID7184964
'Recent studies indicate that propranolol suppresses the normal murine lymphocyte mitogen dose-response(1) and that propranolol has a high affinity for the beta-adrenergic binding site on the murine lymphocyte(2). Since concentrations which suppress the lymphocyte mitogen dose-response are much greater than the Kd for the binding site, it may be suggested ... More
Fluorescently labelled Na+ channels are localized and immobilized to synapses of innervated muscle fibres.
AuthorsAngelides KJ
JournalNature
PubMed ID2422557
'Segregation of voltage-dependent sodium channels to the hillock of motoneurones and nodes of Ranvier in myelinated axons is crucial for conduction of the nerve impulse. Much less is known, however, about the distribution of voltage-dependent Na+ channels on muscle fibres. Recently, Beam et al. have shown that Na+ channels are ... More
Heterogeneous distribution of a basement membrane heparan sulfate proteoglycan in rat tissues.
AuthorsCouchman JR
JournalJ Cell Biol
PubMed ID2959669
'A heparan sulfate proteoglycan (HSPG) synthesized by murine parietal yolk sac (PYS-2) cells has been characterized and purified from culture supernatants. A monospecific polyclonal antiserum was raised against it which showed activity against the HSPG core protein and basement membrane specificity in immunohistochemical studies on frozen tissue sections from many ... More
The cercarial glycocalyx of Schistosoma mansoni.
AuthorsSamuelson JC, Caulfield JP
JournalJ Cell Biol
PubMed ID2985622
'Cercariae, the freshwater stage of Schistosoma mansoni infectious to man, are covered by a single unit membrane and an immunogenic glycocalyx. When cercariae penetrate the host skin, they transform to schistosomula by shedding tails, secreting mucous and enzymes, and forming microvilli over their surface. Here the loss of the glycocalyx ... More
Distribution and lateral mobility of voltage-dependent sodium channels in neurons.
AuthorsAngelides KJ, Elmer LW, Loftus D, Elson E
JournalJ Cell Biol
PubMed ID2454930
'Voltage-dependent sodium channels are distributed nonuniformly over the surface of nerve cells and are localized to morphologically distinct regions. Fluorescent neurotoxin probes specific for the voltage-dependent sodium channel stain the axon hillock 5-10 times more intensely than the cell body and show punctate fluorescence confined to the axon hillock which ... More
Redistribution and shedding of flagellar membrane glycoproteins visualized using an anti-carbohydrate monoclonal antibody and concanavalin A.
AuthorsBloodgood RA, Woodward MP, Salomonsky NL
JournalJ Cell Biol
PubMed ID3009491
'Two carbohydrate-binding probes, the lectin concanavalin A and an anti-carbohydrate monoclonal antibody designated FMG-1, have been used to study the distribution of their respective epitopes on the surface of Chlamydomonas reinhardtii, strain pf-18. Both of these ligands bind uniformly to the external surface of the flagellar membrane and the general ... More
Agglutination of isolated rod outer segments by lectins.
AuthorsBridges CD
JournalInvest Ophthalmol Vis Sci
PubMed ID6969711
'The interactions of ten lectins with rod outer segment (ROS) plasma membranes and the effects of various haptene sugars were examined by studying agglutination in suspensions of washed ROSs. It is concluded that agglutination sites on the ROS membrane involve receptors that are recognized by eight lectins with haptene sugar ... More
A fluorescent lectin array using supramolecular hydrogel for simple detection and pattern profiling for various glycoconjugates.
AuthorsKoshi Y, Nakata E, Yamane H, Hamachi I
JournalJ Am Chem Soc
PubMed ID16895406
'Because sugar and its derivatives play important roles in various biological phenomena, the rapid and high-throughput analysis of various glycoconjugates is keenly desirable. We describe herein the construction of a novel fluorescent lectin array for saccharide detection using a supramolecular hydrogel matrix. In this array, the fluorescent lectins were noncovalently ... More
Lectins facilitate calcium-induced fusion of phospholipid vesicles containing glycosphingolipids.
AuthorsDüzgünes N, Hoekstra D, Hong K, Papahadjopoulos D
JournalFEBS Lett
PubMed ID6745438
'Ca2+-induced fusion of phospholipid vesicles containing globoside (GL-4) or disialoganglioside (GDla) is several-fold slower than the fusion of the pure phospholipid vesicles. Lectins specific for these glycosphingolipids, soybean agglutinin and wheat germ agglutinin, respectively, enhance the rate of fusion when added to the vesicle suspension before the introduction of Ca2+. ... More
Controlled binding of liposomes to cultured cells by means of lectins.
AuthorsLiautard JP, Vidal M, Philippot JR
JournalCell Biol Int Rep
PubMed ID3841030
'Lectin-mediated binding of liposomes to Hela cells was analyzed as a function of different parameters. We show that the amount of lectin covalently bound to liposomes can be accurately controlled. We chose to work with 500 - to 1 000 molecules of WGA bound per liposome of 1 micron diameter. ... 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
Histological, histochemical, and fine structural observations on the spleen of seals.
AuthorsSchumacher U, Welsch U
JournalAm J Anat
PubMed ID3661458
'Spleens of three species of Antarctic seals with different diving habits (Weddell seal, crabeater seal, and fur seal) have been studied with histological, histochemical, and electron microscopic methods. The spleens can be classified as nonsinusoidal, with capsule and trabeculae rich in innervated smooth muscle cells. The trabecular system is particularly ... More
Polar effects of concanavalin A on the plasma membrane/cytoskeleton complex in a molluscan egg.
AuthorsDohmen MR, Speksnijder JE, Teerds KJ
JournalCell Biol Int Rep
PubMed ID6744428
Gel protein stains: glycoproteins.
AuthorsGander JE
JournalMethods Enzymol
PubMed ID6201699
Specific fluorescent analysis of serum glycoproteins following cellulose acetate electrophoresis.
AuthorsVarma RS, Varma R
JournalBiochem Med
PubMed ID6187340
Photoinduced cell killing and crosslinking of fluorescein conjugated concanavalin A to cell surface proteins.
AuthorsLepock JR, Campbell SD, Gruber M, Kruuv J
JournalBiochem Biophys Res Commun
PubMed ID526271
The carbohydrate moiety of rhodopsin: lectin-binding, chemical modification and fluorescence studies.
AuthorsRenthal R, Steinemann A, Stryer L
JournalExp Eye Res
PubMed ID4361169
Cell shape and membrane receptor dynamics. Modulation by the cytoskeleton.
AuthorsAlbertini DF, Herman B
JournalCell Muscle Motil
PubMed ID6322968
Resonance energy transfer microscopy.
AuthorsHerman B
JournalMethods Cell Biol
PubMed ID2648111
Affinity cytochemistry: the localization of lectin and antibody receptors on erythrocytes via the avidin-biotin complex.
AuthorsBayer EA, Wilchek M, Skutelsky E
JournalFEBS Lett
PubMed ID976475
Behavior of mitochondria in the living cell.
AuthorsBereiter-Hahn J
JournalInt Rev Cytol
PubMed ID2246114
Fluorescence microscopy methods for yeast.
AuthorsPringle JR, Preston RA, Adams AE, Stearns T, Drubin DG, Haarer BK, Jones EW
JournalMethods Cell Biol
PubMed ID2476649
Staining of glycoproteins in polyacrylamide and agarose gels with fluorescent lectins.
AuthorsFurlan M, Perret BA, Beck EA
JournalAnal Biochem
PubMed ID91330
Luminescence digital imaging microscopy.
AuthorsJovin TM, Arndt-Jovin DJ
JournalAnnu Rev Biophys Biophys Chem
PubMed ID2660827
Coincidence of endoplasmic reticulum pattern as visualized by FITC-Con A-fluorescence and electron microscopy.
AuthorsFuhrmann C, Bereiter-Hahn J
JournalHistochemistry
PubMed ID6201466
FITC-Con A-fluorescence and electron microscopy have been used in parallel for the visualization of rough endoplasmic reticulum in Xenopus laevis tadpole heart primary tissue culture cells. rER distribution pattern as revealed by both methods is compared by superimposition of images of the same cell. Coincidence of rER distribution is established. ... More
Developmental changes in the lateral diffusion of Leydig cell membranes measured by the FRAP method.
AuthorsZlatanov IV, Foley M, Birmingham J, Garland PB
JournalFEBS Lett
PubMed ID3653400
A simple method for isolation and fluorescence labelling of Leydig cells (L-cells) from rat testes was developed. Lateral diffusion coefficients of both lipid and protein membrane fluorescent probes were measured by the method of fluorescence recovery after photobleaching (FRAP). Age-dependent changes in diffusibility of membrane lipids and proteins were discovered. ... More
Fluorescein-concanavalin A conjugates distinguish between normal and malignant human cells: a preliminary report.
AuthorsLouis CJ, Wyllie RG
JournalExperientia
PubMed ID6788591
A method is described for using a fluorescein isothiocyanate concanavalin A conjugate to stain human cell membranes in formalin fixed paraffin embedded tissue. 57 neoplastic and normal tissue sites were examined. In 54 malignant tumours, bright green fluorescence was confined to the cell membranes while in 23 benign tumours and ... More
Lateral motion of fluorescent molecules embedded into cell membranes of clonal myogenic cells, L6, changes upon cell maturation.
AuthorsKawasaki Y, Wakayama N, Seto-Ohshima A
JournalFEBS Lett
PubMed ID3360141
The lateral motion of fluorescent molecules embedded into cell membranes of myogenic cell line, L6, was measured. The motion of S-F-ConA became faster at cell fusion stage, and became slower after fusion. On the other hand, the motion of lipid analog, F18, was not changed at cell fusion stage. However, ... More
The carbohydrates associated with the surfaces of hamster alveolar macrophages: a study using fluorescein-conjugated lectins.
AuthorsMeban C
JournalJ Anat
PubMed ID3935630
Alveolar macrophages were isolated from the lungs of hamsters by lavage and centrifugation. The harvesting procedure yielded about 5.2 X 10(6) cells per animal and the majority of these cells were viable as assessed by a Trypan blue exclusion test. Monolayers of macrophages were labelled with a series of FITC-conjugated ... More
A technique for fluorescence microscopy in semithin sections.
AuthorsOjeda JL, Ros MA, Icardo JM
JournalStain Technol
PubMed ID2699110
We describe here a procedure to improve contrast and resolution in fluorescence microscopy of sectioned tissues. Tissue fragments were fixed in ethanol-glacial acetic acid, embedded in diethylene glycol distearate, and semithin sectioned. This method maintains tissue antigenicity while preserving the structure of cells and tissues. The thinness of the sections ... More
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
Spatial distribution of cortical proteins in cells of epithelial sheets.
AuthorsOpas M, Kalnins VI
JournalCell Tissue Res
PubMed ID3919953
In the differentiated pigmented epithelial cells of the retina (RPE) of chick embryos cytoskeletal proteins are found in polygonal rings located in the cell cortex. Within the cortical rings of the RPE cells vinculin and spectrin occupy a characteristic position closest to the plasma membrane; actin is found farther away, ... More
Inhibition of polymorphonuclear leukocyte capping by a chemotactic factor.
AuthorsMandell BF, Spilberg I, Lichtman J
JournalJ Immunol
PubMed ID321690
Incubation of human polymorphonuclear leukocytes with colchicine and fluorescein-concanavalin A leads to the formation of a polarized cap of fluorescence not seen if cells are incubated with fluorescein-Con A along. When cells are preincubated with a chemotactic factor before colchicine treatment, the capping is inhibited in a dose-related manner. Studies ... 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
Spot analysis for glycoprotein determination in the nanogram range.
AuthorsNeuhoff V, Ewers E, Huether G
JournalHoppe Seylers Z Physiol Chem
PubMed ID6796489
A flexible method for glycoprotein determination with microliter-volumes using spot analysis ("Tüpfelprobe") on cellulose acetate layers is described. With glucose oxidase as an example of a glycoprotein and fluorescein isothiocyanate-labelled Concanavalin A, a sensitivity of 10 ng is reached; in combination with horseradish peroxidase 1 ng of glucose oxidase can ... More
Flow cytometric analysis of concanavalin A binding to isolated Golgi fractions from rat liver.
AuthorsGuasch RM, Guerri C, O'Connor JE
JournalExp Cell Res
PubMed ID8319765
Flow cytometry (FCM) has been used repeatedly to study lectin binding to whole cells. However, there are very few attempts to analyze glycoconjugates in isolated subcellular organelles. We have applied FCM to quantitate the specific binding of fluorescein-conjugated concanavalin A (FITC-Con A) to isolated cis and trans fractions of rat ... More
Assessment of fluorochromes for two-photon laser scanning microscopy of biofilms.
AuthorsNeu TR, Kuhlicke U, Lawrence JR
JournalAppl Environ Microbiol
PubMed ID11823234
A major limitation for the use of two-proton laser scanning microscopy (2P-LSM) in biofilm and other studies is the lack of a thorough understanding of the excitation-emission responses of potential fluorochromes. In order to use 2P-LSM, the utility of various fluorochromes and probes specific for a range of biofilm constituents ... 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