Indirect immunofluorescence localization of ponticulin in motile cells.
AuthorsWuestehube LJ, Chia CP, Luna EJ
JournalCell Motil Cytoskeleton
PubMed ID2673549
'Ponticulin is the major actin-binding integral glycoprotein in plasma membranes isolated from log-phase Dictyostelium discoideum amebae. As such, this protein appears to be an important link between the plasma membrane and actin filaments (Wuestehube and Luna: Journal of Cell Biology 105:1741-1751, 1987). In this study, indirect immunofluorescence microscopy was used ... More
A functional YNKI motif in the short cytoplasmic tail of varicella-zoster virus glycoprotein gH mediates clathrin-dependent and antibody-independent endocytosis.
AuthorsPasieka TJ, Maresova L, Grose C,
JournalJ Virol
PubMed ID12634377
'The trafficking of varicella-zoster virus (VZV) gH was investigated under both infection and transfection conditions. In initial endocytosis assays performed in infected cells, the three glycoproteins gE, gI, and gB served as positive controls for internalization from the plasma membrane. Subsequently, we discovered that gH in VZV-infected cells was also ... More
Rotavirus NSP4 induces a novel vesicular compartment regulated by calcium and associated with viroplasms.
AuthorsBerkova Z, Crawford SE, Trugnan G, Yoshimori T, Morris AP, Estes MK
JournalJ Virol
PubMed ID16731945
'Rotavirus is a major cause of infantile viral gastroenteritis. Rotavirus nonstructural protein 4 (NSP4) has pleiotropic properties and functions in viral morphogenesis as well as pathogenesis. Recent reports show that the inhibition of NSP4 expression by small interfering RNAs leads to alteration of the production and distribution of other viral ... More
Isolation of bright aggregate fluctuations in a multipopulation image correlation spectroscopy system using intensity subtraction.
AuthorsRocheleau JV, Wiseman PW, Petersen NO
JournalBiophys J
PubMed ID12770905
'Image correlation spectroscopy allows sensitive measurement of the spatial distribution and aggregation state of fluorescent membrane macro molecules. When studying a single population system (i.e., aggregates of similar brightness), an accurate measure can be made of the aggregate number per observation area, but this measurement becomes much more complex in ... More
Microstructural differences between single-species and dual-species biofilms of Streptococcus mutans and Veillonella parvula, before and after exposure to chlorhexidine.
AuthorsKara D, Luppens SB, van Marle J, Ozok R, ten Cate JM,
JournalFEMS Microbiol Lett
PubMed ID17403046
'Dual-species biofilms of Streptococcus mutans and Veillonella parvula are less susceptible to antimicrobials than single-species biofilms of the same microorganisms. The microstructure of single and dual-species biofilms of S. mutans and/or V. parvula was visualized to find out whether the spatial arrangement of bacteria in biofilms is related to survival ... More
Actin coating of secretory granules during regulated exocytosis correlates with the release of rab3D.
'The present study describes a novel phenomenon in pancreatic acinar cells undergoing regulated exocytosis. When acinar cell preparations were challenged with the secretagogue carbamylcholine, a subpopulation of zymogen granules became coated with filamentous actin. These zymogen granules were always in proximity of the acinar cell apical membrane (the site of ... More
Texas Red, a hydrophilic, red-emitting fluorophore for use with fluorescein in dual parameter flow microfluorometric and fluorescence microscopic studies.
AuthorsTitus JA, Haugland R, Sharrow SO, Segal DM
JournalJ Immunol Methods
PubMed ID6806389
The sulfonylchloride derivative of the red-emitting fluorophore, sulforhodamine 101, has been synthesized in order to provide a reagent for coupling to amino groups on proteins and other compounds, and it is now commercially available under the name 'Texas Red'. Texas Red conjugates of antibodies and other proteins have been prepared ... More
Requirement of Rac1 in the development of cardiac hypertrophy.
AuthorsSatoh M, Ogita H, Takeshita K, Mukai Y, Kwiatkowski DJ, Liao JK
JournalProc Natl Acad Sci U S A
PubMed ID16651530
The development of cardiac hypertrophy is mediated, in part, by increase in NADPH oxidase activity and myocardial oxidative stress. The Rho GTPase, Rac, regulates NADPH oxidase activity through interaction with gp91(phox) and p67(phox) (in which "phox" is phagocyte oxidase). However, it is not known which Rac isoform mediates this effect ... More
A subset of human dendritic cells expresses IgA Fc receptor (CD89), which mediates internalization and activation upon cross-linking by IgA complexes.
AuthorsGeissmann F, Launay P, Pasquier B, Lepelletier Y, Leborgne M, Lehuen A, Brousse N, Monteiro RC
JournalJ Immunol
PubMed ID11123311
Immature dendritic cells (DC) sample Ags within nonlymphoid tissues and acquire exogenous proteins/pathogens via scavenger receptors or Ig FcR such as Fc gamma R and Fc epsilon R. IgA is present in a significant proportion among serum Ig and is the main isotype in mucosae, where DC are numerous. We ... More
Delineation among eight major hematopoietic subsets in murine bone marrow using a two-color flow cytometric technique.
AuthorsSchlueter AJ, Bhatia SK, Li X, Tygrett LT, Yamashita Y, de Vries P, Waldschmidt TJ
JournalCytometry
PubMed ID11260597
BACKGROUND: Many methods have been developed specifically for identifying hematopoietic progenitor cells in murine bone marrow, but few methods allow rapid identification of multiple bone marrow populations. We describe a new, simple method for identifying simultaneously eight populations in murine bone marrow with two-color flow cytometry and phenotypically define these ... More
Disruption of disulfide bonds is responsible for impaired secretion in human complement factor H deficiency.
Factor H, a secretory glycoprotein composed of 20 short consensus repeat modules, is an inhibitor of the complement system. Previous studies of inherited factor H deficiency revealed single amino acid substitutions at conserved cysteine residues, on one allele arginine for cysteine 518 (C518R) and on the other tyrosine for cysteine ... More
Intrasarcomere [Ca2+] gradients in ventricular myocytes revealed by high speed digital imaging microscopy.
AuthorsIsenberg G, Etter EF, Wendt-Gallitelli MF, Schiefer A, Carrington WA, Tuft RA, Fay FS
JournalProc Natl Acad Sci U S A
PubMed ID8643589
Cardiac muscle contraction is triggered by a small and brief Ca2+ entry across the t-tubular membranes, which is believed to be locally amplified by release of Ca2+ from the adjacent junctional sarcoplasmic reticulum (SR). As Ca2+ diffusion is thought to be markedly attenuated in cells, it has been predicted that ... More
Subtype-specific translocation of diacylglycerol kinase alpha and gamma and its correlation with protein kinase C.
AuthorsShirai Y, Segawa S, Kuriyama M, Goto K, Sakai N, Saito N
JournalJ Biol Chem
PubMed ID10827086
We examined the translocation of diacylglycerol kinase (DGK) alpha and gamma fused with green fluorescent protein in living Chinese hamster ovary K1 cells (CHO-K1) and investigated temporal and spatial correlations between DGK and protein kinase C (PKC) when both kinases are overexpressed. DGKalpha and gamma were present throughout the cytoplasm ... More
A high-resolution, fluorescence-based method for localization of endogenous alkaline phosphatase activity.
AuthorsCox WG, Singer VL
JournalJ Histochem Cytochem
PubMed ID10544217
We describe a high-resolution, fluorescence-based method for localizing endogenous alkaline phosphatase in tissues and cultured cells. This method utilizes ELF (Enzyme-Labeled Fluorescence)-97 phosphate, which yields an intensely fluorescent yellow-green precipitate at the site of enzymatic activity. We compared zebrafish intestine, ovary, and kidney cryosections stained for endogenous alkaline phosphatase using ... More
Voltage sensing by fluorescence resonance energy transfer in single cells.
AuthorsGonzález JE, Tsien RY
JournalBiophys J
PubMed ID8534797
A new mechanism has been developed for achieving fast ratiometric voltage-sensitive fluorescence changes in single cells using fluorescence resonance energy transfer. The mechanism is based on hydrophobic fluorescent anions that rapidly redistribute from one face of the plasma membrane to the other according to the Nernst equation. A voltage-sensitive fluorescent ... More
Sphingosine-1-phosphate phosphohydrolase in regulation of sphingolipid metabolism and apoptosis.
AuthorsLe Stunff H, Galve-Roperh I, Peterson C, Milstien S, Spiegel S
JournalJ Cell Biol
PubMed ID12235122
Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid metabolite that regulates diverse biological processes by binding to a family of G protein-coupled receptors or as an intracellular second messenger. Mammalian S1P phosphatase (SPP-1), which degrades S1P to terminate its actions, was recently cloned based on homology to a lipid phosphohydrolase that regulates ... More
Cytoplasmic dynein contains a family of differentially expressed light chains.
AuthorsKing SM, Barbarese E, Dillman JF, Benashski SE, Do KT, Patel-King RS, Pfister KK
JournalBiochemistry
PubMed ID9790665
Cytoplasmic dynein contains a series of accessory proteins associated with the motor containing heavy chains.1 These include three distinct classes of light chains (Mr < approximately 22 000). Here we demonstrate that a previously cloned protein termed rp3 is a bona fide Mr 14 000 light chain component of this ... More
Cellular responses to excess phospholipid.
AuthorsBaburina I, Jackowski S
JournalJ Biol Chem
PubMed ID10092620
Phosphatidylcholine (PtdCho) is the major membrane phospholipid in mammalian cells, and its synthesis is controlled by the activity of CDP:phosphocholine cytidylyltransferase (CCT). Enforced CCT expression accelerated the rate of PtdCho synthesis. However, the amount of cellular PtdCho did not increase as a result of the turnover of both the choline ... More
Distinct effects of fatty acids on translocation of gamma- and epsilon-subspecies of protein kinase C.
AuthorsShirai Y, Kashiwagi K, Yagi K, Sakai N, Saito N
JournalJ Cell Biol
PubMed ID9786959
Effects of fatty acids on translocation of the gamma- and epsilon-subspecies of protein kinase C (PKC) in living cells were investigated using their proteins fused with green fluorescent protein (GFP). gamma-PKC-GFP and epsilon-PKC-GFP predominated in the cytoplasm, but only a small amount of gamma-PKC-GFP was found in the nucleus. Except ... More
Characterization and immunolocalization of bovine N-acetylglucosamine-1-phosphodiester alpha-N-acetylglucosaminidase.
AuthorsMullis KG, Kornfeld RH
JournalJ Biol Chem
PubMed ID8294421
N-Acetylglucosamine-1-phosphodiester alpha-N-acetylglucosaminidase (phosphodiester alpha-GlcNAcase) has been purified 3,000-fold from bovine liver and its kinetic properties determined as described in the previous report (Mullis, K. G., Huynh, M., and Kornfeld, R. (1993) J. Biol. Chem. 269, 1718-1726). This report describes the hydrodynamic and lectin binding properties of phosphodiester alpha-GlcNAcase as well ... More
Distribution of gap junctions in dog and rat ventricle studied with a double-label technique.
AuthorsDolber PC, Beyer EC, Junker JL, Spach MS
JournalJ Mol Cell Cardiol
PubMed ID1338112
To assess the distribution of gap junctions in relation to the cardiac myocyte surface in paraffin sections of dog and rat ventricle, the sarcolemma was labeled with wheat germ agglutinin (WGA1) and gap junctions were labeled with antibodies to cardiac muscle gap junction protein connexin43. WGA labeled all of the ... More
Dynamic measurement of the pH of the Golgi complex in living cells using retrograde transport of the verotoxin receptor.
AuthorsKim JH, Lingwood CA, Williams DB, Furuya W, Manolson MF, Grinstein S
JournalJ Cell Biol
PubMed ID8830769
The B subunit of verotoxin (VT1B) from enterohemorrhagic Escherichia coli is responsible for the attachment of the holotoxin to the cell surface, by binding to the glycolipid, globotriaosyl ceramide. After receptor-mediated endocytosis, the toxin is targeted to the Golgi complex by a process of retrograde transport. We took advantage of ... More
Permeability and gating properties of human connexins 26 and 30 expressed in HeLa cells.
AuthorsBeltramello M, Bicego M, Piazza V, Ciubotaru CD, Mammano F, D'Andrea P
JournalBiochem Biophys Res Commun
PubMed ID12767933
Human connexins 26 and 30 were expressed either through the bicistronic pIRES-EGFP expression vector or as EYFP-tagged chimeras. When transiently transfected in communication-incompetent HeLa cells, hCx26-pIRES transfectants were permeable to dyes up to 622 Da, but were significantly less permeable to 759 Da molecules. Under the same conditions, permeability of ... More