Exocytotic insertion of calcium channels constrains compensatory endocytosis to sites of exocytosis.
AuthorsSmith RM, Baibakov B, Ikebuchi Y, White BH, Lambert NA, Kaczmarek LK, Vogel SS
JournalJ Cell Biol
PubMed ID10684256
'Proteins inserted into the cell surface by exocytosis are thought to be retrieved by compensatory endocytosis, suggesting that retrieval requires granule proteins. In sea urchin eggs, calcium influx through P-type calcium channels is required for retrieval, and the large size of sea urchin secretory granules permits the direct observation of ... More
Identification of a novel domain in two mammalian inositol-polyphosphate 5-phosphatases that mediates membrane ruffle localization. The inositol 5-phosphatase skip localizes to the endoplasmic reticulum and translocates to membrane ruffles following epidermal growth factor stimulation.
AuthorsGurung R, Tan A, Ooms LM, McGrath MJ, Huysmans RD, Munday AD, Prescott M, Whisstock JC, Mitchell CA
JournalJ Biol Chem
PubMed ID12536145
'SKIP (skeletal muscle and kidney enriched inositol phosphatase) is a recently identified phosphatidylinositol 3,4,5-trisphosphate- and phosphatidylinositol 4,5-bisphosphate-specific 5-phosphatase. In this study, we investigated the intracellular localization of SKIP. Indirect immunofluorescence and subcellular fractionation showed that, in serum-starved cells, both endogenous and recombinant SKIP colocalized with markers of the endoplasmic reticulum ... More
The N-terminus of retinol dehydrogenase type 1 signals cytosolic orientation in the microsomal membrane.
AuthorsWang J, Bongianni JK, Napoli JL
JournalBiochemistry
PubMed ID11601977
'We determined the orientation of the SDR (short-chain dehydrogenase/reductase) rat RoDH1 (retinol dehydrogenase type 1) in the endoplasmic reticulum to provide insight into its function in retinol metabolism, and to resolve whether retinoid-metabolizing SDRs differ from several other SDRs by requiring a C-terminal segment for the membrane orientation. In contrast ... More
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
Functional expression, quantification and cellular localization of the Hxt2 hexose transporter of Saccharomyces cerevisiae tagged with the green fluorescent protein.
AuthorsKruckeberg AL, Ye L, Berden JA, van Dam K
JournalBiochem J
PubMed ID10191260
'The Hxt2 glucose transport protein of Saccharomyces cerevisiae was genetically fused at its C-terminus with the green fluorescent protein (GFP). The Hxt2-GFP fusion protein is a functional hexose transporter: it restored growth on glucose to a strain bearing null mutations in the hexose transporter genes GAL2 and HXT1 to HXT7. ... 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
Carbohydrate-labeled fluorescent microparticles and their binding to lectins.
AuthorsKelly TL, Lam MC, Wolf MO
JournalBioconjug Chem
PubMed ID16704193
The preparation of micrometer-sized, cross-linked poly(p-phenyleneethynylene) (PPE) beads to which simple monosaccharides are attached is reported. Mannose, glucose, and galactose derivatives have been synthesized. The fluorescence properties, size distribution, and morphology of these microparticles have been elucidated through fluorimetry, fluorescence confocal microscopy, and scanning electron microscopy. Protein binding assays were ... More
Differential partitioning of pulmonary surfactant protein SP-A into regions of monolayers of dipalmitoylphosphatidylcholine and dipalmitoylphosphatidylcholine/dipalmitoylphosphatidylglycerol.
AuthorsRuano ML, Nag K, Worthman LA, Casals C, Pérez-Gil J, Keough KM
JournalBiophys J
PubMed ID9512012
The interaction of the pulmonary surfactant protein SP-A fluorescently labeled with Texas Red (TR-SP-A) with monolayers of dipalmitoylphosphatidylcholine (DPPC) and DPPC/dipalmitoylphosphatidylglycerol 7:3 w/w has been investigated. The monolayers were spread on aqueous subphases containing TR-SP-A. TR-SP-A interacted with the monolayers of DPPC to accumulate at the boundary regions between liquid ... More
Spatially regulated recruitment of clathrin to the plasma membrane during capping and cell translocation.
AuthorsDamer CK, O'Halloran TJ
JournalMol Biol Cell
PubMed ID10848635
Clathrin-coated vesicles bud from selected cellular membranes to traffic-specific intracellular proteins. To study the dynamic properties of clathrin-coated membranes, we expressed clathrin heavy chain tagged with green fluorescent protein (GFP) in Dictyostelium cells. GFP-clathrin was functional and retained the native properties of clathrin: the chimeric protein formed classic clathrin lattices ... More
ERp29 is a ubiquitous resident of the endoplasmic reticulum with a distinct role in secretory protein production.
AuthorsShnyder SD, Hubbard MJ
JournalJ Histochem Cytochem
PubMed ID11897809
ERp29 was recently characterized biochemically as a novel protein that resides in mammalian endoplasmic reticulum (ER). Here we applied immunochemical procedures at the cellular level to investigate the hypothesized role of ERp29 in secretory protein production. ERp29 was localized exclusively to the ER/nuclear envelope of MDCK cells using confocal immunocytochemistry ... More
Single particle tracking of surface receptor movement during cell division.
AuthorsWang YL, Silverman JD, Cao LG
JournalJ Cell Biol
PubMed ID7962078
We have used fluorescent latex beads to label membrane receptors on cultured NRK cells. Movement of individual beads during cell division was recorded with digital imaging techniques. Surface-bound beads showed no organized movement during metaphase but started to migrate toward the equator approximately 1 min after anaphase onset, when chromosomes ... 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
Transport of bacterial lipopolysaccharide to the golgi apparatus.
AuthorsThieblemont N, Wright SD
JournalJ Exp Med
PubMed ID10449523
Addition of lipopolysaccharide (LPS) to cells in the form of LPS-soluble (s)CD14 complexes induces strong cellular responses. During this process, LPS is delivered from sCD14 to the plasma membrane, and the cell-associated LPS is then rapidly transported to an intracellular site. This transport appears to be important for certain cellular ... More
Bhalin, an Essential Cytoskeleton-Associated Protein of Trypanosoma brucei Linking TbBILBO1 of the Flagellar Pocket Collar with the Hook Complex.