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Topic: Phalloidin


  
  Biophysical Journal: Position and Orientation of Phalloidin in F-Actin Determined by X-Ray Fiber Diffraction Analysis
To determine the position of bound phalloidin experimentally, x-ray fiber diffraction data were obtained from well-oriented sols of F-actin and the phalloidin-F-actin complex.
The position of rhodamine attached to phalloidin in the rhodamine-phalloidin-F-actin complex was also determined, in which the conjugated Leu(OH)^sup 7^ residue was found to face the outside of the filament.
The aim of this study is to discuss one of the key mechanisms that stabilize F-actin by determining the position of bound phalloidin and analyzing its binding interactions to actin subunits.
www.findarticles.com /p/articles/mi_qa3938/is_200504/ai_n13602124   (1463 words)

  
 Phalloidin
Kinosian, H. J., Seldon, L. A., Estes, J. and Gershman, L. (1993) Actin filament annealing in the presence of ATP and phalloidin.
Wendel, H. and Dancker, P. (1986) Kinetics of actin polymerization: influence of ions, temperature, age of F-actin, cytochalsin B and phalloidin.
Wendel, H. and Dancker, P. (1987) Influence of phalloidin on both the nucleation and the elongation phase of actin polymerization.
www.bms.ed.ac.uk /research/others/smaciver/Encyclop/ABP-P/phalloidin.htm   (1158 words)

  
 Actin
Phalloidin binding to actin has been shown to delay the release of inorganic phosphate after ATP hydrolysis (Dancker & Hess, Biochim.
Phalloidin, whose position in the F-actin filament structure is known, would in our model block the exit of the back door pathway and, thus, delay the release of the phosphate.
ATP and the Mg++ ion are represented as white spheres in the center of the protein, which is rendered as a transparent ribbon.
www.ks.uiuc.edu /Research/cell_motility/actin   (510 words)

  
 Nikon MicroscopyU: Fluorescence Microscopy Digital Image Gallery - Indian Muntjac Deer Skin Fibroblast Cells
The culture was fixed and subsequently labeled with DAPI and Alexa Fluor 488 conjugated to phalloidin, targeting DNA in the cell nucleus and the actin cytoskeletal framework, respectively.
Visualizing the Golgi Complex in Deer Skin Cells - A monolayer culture of Indian Muntjac cells was fixed, permeabilized, and blocked with 10-percent normal goat serum in phosphate-buffered saline prior to immunofluorescent labeling with rabbit primary antibodies to giantin, a protein resident in the Golgi complex of mammalian cells.
Alexa Fluor 488 conjugated to phalloidin and DAPI were simultaneously used to counterstain the culture, targeting cytoskeletal F-actin and nuclear DNA, respectively.
www.microscopyu.com /galleries/fluorescence/cells/muntjac/muntjac.html   (1491 words)

  
 Exercise #3 - Effect of Cytochalasin D on Microfilament Organization and
By careful inspection of your results, you should be able to describe the effects of cytochalasin D on the organization of the different actin containing structures in a fibroblastic cell.
Phalloidin, an alkaloid compound produced by the mushroom Amanita phalloides, binds to F-actin and prevents depolymerization.
Thus, phalloidin conjugated with a reactive form of fluorescein, fluorescein isothiocyanate (FITC), is a convenient molecular probe for F-actin.
www.unr.edu /biology/Biol395_S_05/Lab3.htm   (1814 words)

  
 Technical Report 99-006: Cell Biological Applications of Fluorescence Microscopy: Techniques for Staining Specific ...
Phalloidin selectively binds and stabilizes polymerized, filamentous actin without binding monomeric actin and its non-specific staining is negligible.
Phalloidin is usually purchased as a lyophilized powder and is reconstituted in methanol or DMSO.
Phalloidin is cell-impermeable, so specimens to be stained must be fixed and detergent-permeabilized.
www.itg.uiuc.edu /publications/techreports/99-006/actin.htm   (297 words)

  
 Olympus Microscopy Resource Center: Specialized Microscopy Techniques - Fluorescence Digital Image Gallery - Indian ...
Visualizing the Golgi Complex with Immunofluorescence in Indian Muntjac Fibroblast Cell Cultures - A log phase monolayer culture of Indian Muntjac cells was fixed, permeabilized, and blocked with 10-percent normal goat serum in phosphate-buffered saline prior to immunofluorescent labeling with rabbit primary antibodies to giantin, a protein resident in the Golgi complex of mammalian cells.
The Golgi Apparatus in Indian Muntjac Cells - A culture of Indian Muntjac fibroblasts was fixed, permeabilized, and blocked with 10-percent normal goat serum in phosphate-buffered saline prior to immunofluorescent labeling with primary antibodies to giantin, a protein resident in the Golgi complex of mammalian cells.
The cells were subsequently labeled with the conjugated phalloidin and counterstained with the bisbenzimide reagent.
www.olympusmicro.com /primer/techniques/fluorescence/gallery/cells/muntjac/muntjaccells.html   (2008 words)

  
 Jahrbuch-CD der MPG 2003 - Growth inhibition and changes i
Effects on morphology and microfilament structure caused by phalloidin, phallacidin, and some semisynthetic phalloidin derivatives were studied in vegetative cells of the green alga Acetabularia acetabulum (L.) Silva.
All phalloidin derivatives (except for phalloidin itself) caused growth stop of the alga after 1 day and (except for the fluorescein-labeled phalloidin) death of the cells after 4-7 days.
Indeed, the most lipophilic phalloidin derivatives benzoylphalloidin and dithiolanophalloidin were the most active in causing growth stop at ca.
www.mpg.de /forschungsergebnisse/wissVeroeffentlichungen/archivListenJahrbuch/2003/66/publZIM20.html   (216 words)

  
 Olympus FluoView Resource Center: Confocal Gallery - Indian Muntjac Deer Skin Fibroblast Cells
The cells were subsequently labeled with the conjugated phalloidin and counterstained with the SYTOX reagent.
The cells were subsequently labeled with the conjugated phalloidin and counterstained with the cyanine monomer (TO-PRO-3) reagent.
The cells were subsequently labeled with the conjugated phalloidin and counterstained with the anthraquinone (DRAQ5) reagent.
www.olympusfluoview.com /gallery/cells/muntjac/muntjaccells.html   (1218 words)

  
 Phalloidin-Eosin Followed by Photo-oxidation: A Novel Method for Localizing F-Actin at the Light and Electron ...
Light microscopic images of phalloidin labeling in regions of rat brain with high concentrations of dendritic spines.
Electron microscopic analysis shows that phalloidin labeling seen at the light microscopic level is concentrated in postsynaptic structures (arrow) and not in the presynaptic mossy fiber terminal (MF).
Cooper JA (1987) Effects of cytochalasin and phalloidin on actin.
www.jhc.org /cgi/content/full/49/11/1351   (4141 words)

  
 Olympus Microscopy Resource Center: Specialized Microscopy Techniques - Fluorescence Digital Image Gallery - ...
The culture was subsequently stained with Alexa Fluor 568 conjugated to phalloidin to reveal details of the filamentous actin network, and DAPI for DNA in the nucleus.
After fixation and permeabilization, the cells were counterstained for filamentous actin with Alexa Fluor 546 conjugated to phalloidin and for DNA with the nuclear-specific dye, DAPI.
The cells were subsequently counterstained with Alexa Fluor 568 conjugated to phalloidin to localize the filamentous actin network, and the nucleic acid stain DAPI to label DNA in the nucleus.
www.olympusmicro.com /primer/techniques/fluorescence/gallery/cells/mdck/mdckcells.html   (1799 words)

  
 Actin: Dissecting the Structural Basis of its Oligomerization, Polymerization and Polymorphism
As phalloidin does not bind to monomeric actin, it appears not to significantly acti vate G-actin under low-salt conditions.
However, phalloidin tightly binds to and thereby strongly stabilizes nuclei and growing filaments (i.e., by a 'locking-in' mechanism) so that slowly but defi nitely the net equilibrium is shifted toward F-actin filament formation.
Moreover, we have systematically investigated the effect of the divalent cation (i.e., Mg 2+, Ca2+, or none) bound to the HAS of the G-actin molecule in terms of the mechanical properties and 3D structure of the resulting F-actin filaments (i.e., polymerized in the presence of 100mM KCl).
www.scripps.edu /~stoffler/publ/PDF/BioBull98/BioBull98.html   (1541 words)

  
 cGMP-dependent ADP depolymerization of actin mediates estrogen increase in cervical epithelial permeability -- ...
B: filters were washed and reincubated in modified Ringer solution in the absence (C, control) or presence of one of the indicated concentrations of phalloidin for 60 min.
Kinetics and thermodynamics of phalloidin binding to actin filaments from three divergent species.
Effects of cytochalasin, phalloidin and pH on the elongation of actin.
ajpcell.physiology.org /cgi/content/full/279/6/C2028   (5458 words)

  
 Phosphatidylinositol 3-Kinase Regulates Nuclear Translocation of NF-E2-Related Factor 2 through Actin Rearrangement in ...
Phalloidin is an agent that prevents actin filaments from depolymerization (Lader et al., 1999
Inhibition of t-BHQ-inducible Nrf2 translocation and rGSTA2 induction by phalloidin.
C, the effect of phalloidin on the levels of nuclear actin coimmunoprecipiated with Nrf2.
molpharm.aspetjournals.org /cgi/content/full/62/5/1001   (6385 words)

  
 Photo Gallery
Cells are labeled with rhodamine phalloidin (left) and on the right are complimentary pairs of barrel cacti for comparison.
An endothelial cell expressing dominant negative PAK (green), stained for focal adhesions with anti-vinculin (yellow-green) and for actin stress fibers with rhodamine phalloidin (red).
An endothelial cell triple labeled (on the right) with rhodamine phalloidin (red), anti-Beta 3 (blue) and wow-1 (green), and also showing a ratio image (darker green) and the USA.
www.scripps.edu /cb/schwartz/section4.htm   (386 words)

  
 Mechanical Fluctuations of the Membrane-Skeleton Are Dependent on F-Actin ATPase in Human Erythrocytes -- Tuvia et al. ...
The effect of phalloidin or DNase I on amplitude distribution of CMF in RBC ghosts, which were perfused with MgATP.
Phalloidin, is known to specifically and tightly bind to F-actin (De La Cruz and Pollard, 1994
The effect of phalloidin on the maximal amplitude of CMF in RBC ghosts in the presence of 1 mM MgATP.
www.jcb.org /cgi/content/full/141/7/1551   (7113 words)

  
 Tracking Retrograde Flow in Keratocytes: News from the Front -- Vallotton et al. 16 (3): 1223 -- Molecular Biology of ...
Continuous staining of total F-actin in C reveals high actin density at the front of the lamellipodium, indicating that actin distribution was not affected by injection of phalloidin.
Green arrows in kymographs indicate the forward movement of the cell edge, red arrows indicate the retrograde flow of the lamellipodium network, and yellow arrow indicates the anterograde flow of phalloidin speckles in the cell body.
The keratocyte was injected with a low concentration of rhodamine phalloidin and imaged in the two modes with a 2-s interval between fluorescence and phase contrast images.
www.molbiolcell.org /cgi/content/full/16/3/1223   (5725 words)

  
 Effects of the taxanes paclitaxel and docetaxel on edema formation and interstitial fluid pressure -- Brønstad ...
Phalloidin (125 µg) in saline (0.25 ml) was given intravenously
with phalloidin, the subsequent increase in the amount/level
Effect of the cytoskeletal fixation agent phalloidin on transcapillary albumin transport and interstitial fluid pressure in anaphylaxis in the wistar rat.
ajpheart.physiology.org /cgi/content/full/287/2/H963   (3454 words)

  
 Cytoskeleton modulates coupling between availability and activation of cardiac sodium channel -- Maltsev and Undrovinas ...
We used Cyto-D as disrupter and phalloidin as stabilizer of an F-actin-based cytoskeleton.
Phalloidin produced an apparent uncoupling of availability and activation observed as asynchronous shifts in the time course
In the presence of phalloidin, the effect of taxol was
ajpheart.physiology.org /cgi/content/full/273/4/H1832   (4325 words)

  
 Phalloidin Binding and Rheological Differences among Actin Isoforms
Actin is a highly conserved protein in eukaryotes, yet different isoforms of this protein can be found within the same cell.
To begin to explore whether isoactin sequence diversity leads to functional differences in actin filaments, we have examined the phalloidin binding kinetics and the bulk rheologic properties of purified actin isoforms from a variety of eukaryotic sources.
Phalloidin dissociates from all mammalian actin isoforms tested at the same slow rate, while dissociation from yeast actin is 1 order of magnitude more rapid.
pubs.acs.org /cgi-bin/abstract.cgi/bichaw/1996/35/i45/abs/bi961326g.html   (171 words)

  
 Chlamydia pneumoniae Affect Surfactant Trafficking and Secretion Due to Changes of Type II Cell Cytoskeleton -- Wissel ...
Preincubation without and with phalloidin was performed for 30 min and cells were infected or not infected thereafter for 3 h with Cpn.
Effect of phalloidin and paclitaxel of intracellular pathway of Cpn in type II cells.
(D) With phalloidin preincubated type II cells and for 12 h with Cpn infected were labeled with Cpn and then labeled with primary mouse monoclonal M6PR antibodies and Cy3-conjugated Fab fragments goat anti-mouse IgG (red).
ajrcmb.atsjournals.org /cgi/content/full/29/3/303   (6085 words)

  
 A Correlative Analysis of Actin Filament Assembly, Structure, and Dynamics -- Steinmetz et al. 138 (3): 559 -- The ...
in the absence and presence of the mushroom toxin phalloidin.
in the absence of phalloidin (a), and in the presence of a 2:1
and phalloidin on the overall course of polymerization.
www.jcb.org /cgi/content/full/138/3/559   (8205 words)

  
 Changes in Orientation of Actin during Contraction of Muscle -- Borejdo et al. 86 (4): 2308 -- Biophysical Journal
phalloidin bound to actin between 3 and 4 s.
Phalloidin unzips nebulin from thin filaments in skeletal myofibrils.
Studies on conformation of F-actin in muscle fibers in the relaxed state, rigor, and during contraction using fluorescent phalloidin.
www.biophysj.org /cgi/content/full/86/4/2308   (4807 words)

  
 Impaired recycling of synaptic vesicles after acute perturbation of the presynaptic actin cytoskeleton -- Shupliakov et ...
The images were obtained from two different preparations in axons injected with similar amounts of Oregon green-coupled phalloidin.
Phalloidin Identifies a Dynamic Actin Cytomatrix in the Endocytic Zone.
Synaptic vesicles (open arrows) are tethered along filaments (thin arrows) extending from the endocytic zone toward the margin of the vesicle cluster.
www.pnas.org /cgi/content/full/99/22/14476   (3909 words)

  
 Phalloidin   (Site not responding. Last check: 2007-09-10)
Phalloidin is a mushroom poison that binds directly to actin, the principle protein of cytoplasmic microfilaments.
By conjugating phalloidin to the fluorescent dye rhodamine, one can observe the location of microfilaments in fixed cells by use of the fluorescence microscope.
Put the waste phalloidin into the labeled waste container.
www.gettysburg.edu /~rsorense/Celllab04/phalloidin.htm   (591 words)

  
 Photo Gallery -- Cellular Imaging
This cell was fixed and stained with phalloidin to reveal the F-actin distribution (GREEN), tubulin antibodies to reveal the microtubule distribution (RED), and Dapi to show the location of the nucleus (BLUE).
This cell was stained with phalloidin to reveal the F-actin distribution (RED), DAPI to show the location of the nucleus (YELLOW).
Neurons were treated with CdCl for 30 minutes prior to fixation and stained with phalloidin and DNAseI to reveal the F-actin (RED) and g-actin distribution.
www.canisius.edu /biology/cell_imaging/gallery.asp   (390 words)

  
 A Role of Actin Filament in Synaptic Transmission and Long-Term Potentiation -- Kim and Lisman 19 (11): 4314 -- Journal ...
The effect of phalloidin was measured as the ratio of the average EPSC with phalloidin application over that with DMSO alone.
The larger decay in the ratio was produced by phalloidin (filled circles; n = 9; from B) compared with that with DMSO control (open circles; n = 8; from A), which indicates a more selective effect of phalloidin on LTP maintenance.
Collingridge GL, Kehl SJ, McLennan H (1983) Excitatory amino acids in synaptic transmission in the Schaffer collateral-commissural pathway of the rat hippocampus.
www.jneurosci.org /cgi/content/full/19/11/4314   (7521 words)

  
 Alterations in the Actin Cytoskeleton of Pollen Tubes Are Induced by the Self-Incompatibility Reaction in Papaver ...
A distinct, apparently continuous layer of phalloidin labeling is visible adjacent to the plasma membrane.
Almost all phalloidin labeling is present in the form of large punctate foci.
(C) The line scan demonstrates that the amount of phalloidin labeling of F-actin in the cortical region is approximately fivefold that in the lumen of the pollen tube.
www.plantcell.org /cgi/content/full/12/7/1239   (7410 words)

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