Imaging the Lateral Distribution of Fluorescently Labeled Membrane Components of Human Erythrocytes Under Deformation

1999 ◽  
Vol 5 (S2) ◽  
pp. 1044-1045
Author(s):  
D.W. Knowles ◽  
N. Mohandas ◽  
C. Ortiz de Solorzano ◽  
S.J. Lockett
Keyword(s):  
Erythrocyte Membrane ◽  
Cell Biology ◽  
Fluorescence Labeling ◽  
Dimensional Structure ◽  
Biological Macromolecules ◽  
Membrane Component ◽  
Cytoskeletal Network ◽  
Membrane Rigidity ◽  
Semi Solid ◽  
Membrane Components

The human erythrocyte membrane comprises many different biological macromolecules arranged into a cohesive two dimensional structure. It has long been a model system for studying cell membrane component interactions and their related underlying cell biology. Erythrocyte membrane rigidity emanates from its hexagonal semi-solid cytoskeletal network of spectrin polymers joined at junctional complexes by globular proteins. The network supports a 2D fluid double layer of lipid in which is dissolved a large array of receptor proteins some of which completely span the lipid bilayer and link to the underlying cytoskeletal network.To study membrane component - component interactions, we have used the technique of fluorescence imaged microdeformation (1). This technique combines fluorescence labeling of specific membrane components, single cell microdeformation and state-of-the-art image collection and analysis to map the distribution of labeled components on the surface of the cell.

Distributed Modal Voltages of Nonlinear Paraboloidal Shells With Distributed Neurons

2004 ◽  
Vol 126 (1) ◽  
pp. 47-53 ◽  
Author(s):  
H. S. Tzou ◽  
J. H. Ding
Keyword(s):  
Dynamic Responses ◽  
Dynamic State ◽  
Structural Components ◽  
Shells Of Revolution ◽  
Membrane Component ◽  
Fully Integrated ◽  
In Situ Sensors ◽  
Membrane Components ◽  
Paraboloidal Shell

Effective health monitoring and distributed control of advanced structures depends on accurate measurements of dynamic responses of elastic structures. Conventional sensors used for structural measurement are usually add-on “discrete” devices. Lightweight distributed thin-film piezoelectric neurons fully integrated (laminated or embedded) with structural components can serve as in-situ sensors monitoring structure’s dynamic state and health status. This study is to investigate modal voltages and detailed signal contributions of linear or nonlinear paraboloidal shells of revolution laminated with piezoelectric neurons. Signal generation of distributed neuron sensors laminated on paraboloidal shells is defined first, based on the open-voltage assumption and Maxwell’s principle. The neuron signal of a linear paraboloidal shell is composed of a linear membrane component and a linear bending component; the signal of a nonlinear paraboloidal shell is composed of nonlinear and linear membrane components and a linear bending component due to the von Karman geometric nonlinearity. Signal components and distributed modal voltages of linear and nonlinear paraboloidal shells with various curvatures and thickness are investigated.

scholarly journals Human erythrocytes adhering to schistosomula of Schistosoma mansoni lyse and fail to transfer membrane components to the parasite.

1985 ◽  
Vol 101 (1) ◽  
pp. 158-166 ◽  
Author(s):  
J P Caulfield ◽  
C M Cianci
Keyword(s):  
Schistosoma Mansoni ◽  
Erythrocyte Membrane ◽  
Plasma Membranes ◽  
Freeze Fracture ◽  
Human Erythrocytes ◽  
Transmission Microscopy ◽  
Carbocyanine Dyes ◽  
Host Membrane ◽  
Membrane Components ◽  
20 Nm

We studied the adherence of human erythrocytes to larvae of the intravascular parasite Schistosoma mansoni by transmission microscopy, freeze fracture, and fluorescence techniques. In addition, we used the adherent cells to investigate the problem of host antigen acquisition. Schistosomula were cultured for from 24 to 48 h after transformation in order to clear the remnants of the cercarial glycocalyx. In some cases, the worms were preincubated with wheat germ agglutinin to promote adherence of the erythrocytes. The results were similar with and without the lectin except that more cells attached to the lectin-coated parasites. Erythrocytes adhered within a few hours and, unlike neutrophils, did not fuse with the parasite. A layer of 10-20-nm electron dense material separated the outer leaflets of the tegumental and plasma membranes. In addition, many deformed and lysed cells were seen on the parasite surface. The ability of the worm to acquire erythrocyte membrane constituents was tested with carbocyanine dyes, fluorescein covalently conjugated to glycophorin, monoclonal antibodies against B and H blood group glycolipids, and rabbit alpha-human erythrocyte IgG. In summary, glycophorin, erythrocyte proteins, and glycolipids were not transferred to the parasite membrane within 48 h. Carbocyanine dyes were rapidly transferred to the parasite with or without lectin preincubation. Thus, the dye in the worm membrane came from both adherent and nonadherent cells. These studies suggest that, in the absence of membrane fusion, the parasite may acquire some lipid molecules similar in structure to host membrane glycolipids by simple transfer through the medium but that B and H glycolipids and erythrocyte membrane proteins are not transferred from adhering cells to the worm.

Biomolecular dynamics: A report from a workshop in Gysinge, Sweden, October 4–7, 1982

1984 ◽  
Vol 17 (2) ◽  
pp. 125-151 ◽  
Author(s):  
Olle Edholm ◽  
Lennart Nilsson ◽  
Otto Berg ◽  
Måns Ehrenberg ◽  
Flora Claesens ◽  
...  
Keyword(s):  
Waller Factor ◽  
Biological Macromolecules ◽  
Time Resolved ◽  
X Ray Crystallography ◽  
Biomolecular Dynamics ◽  
Debye Waller Factor ◽  
Membrane Components ◽  
Theoretical Simulations ◽  
Structure Of Proteins ◽  
Molecular Picture

From the results of X-ray crystallography a wealth of information is now available concerning the detailed molecular structure of proteins, nucleic acids, and membrane components. This has made it possible to apply successfully various spectroscopie techniques for time resolved studies as well as theoretical simulations of internal molecular dynamics in the biological macromolecules and molecular aggregates. We were particularly pleased to see professor Ivar Waller among the participants of the workshop since new use of the wellknown Debye–Waller factor has greatly contributed to this development. A molecular picture is presently emerging including the dimension of time which ultimately will give us a detailed understanding of the functional interactions between biomolecules in general, and in particular enzyme catalysis, nucleic acid functions, and transport of matter and information through membranes.

scholarly journals Unique inducible filamentous motility identified in pathogenic Bacillus cereus group species

2020 ◽  
Vol 14 (12) ◽  
pp. 2997-3010
Author(s):  
Martha M. Liu ◽  
Shannon Coleman ◽  
Lauren Wilkinson ◽  
Maren L. Smith ◽  
Thomas Hoang ◽  
...  
Keyword(s):  
Bacterial Biomass ◽  
Solid Surfaces ◽  
Ecological Significance ◽  
Rna Seq ◽  
Bacillus Cereus Group ◽  
Control Rod ◽  
Surface Motility ◽  
Semi Solid ◽  
Membrane Components ◽  
Group Species

Abstract Active migration across semi-solid surfaces is important for bacterial success by facilitating colonization of unoccupied niches and is often associated with altered virulence and antibiotic resistance profiles. We isolated an atmospheric contaminant, subsequently identified as a new strain of Bacillus mobilis, which showed a unique, robust, rapid, and inducible filamentous surface motility. This flagella-independent migration was characterized by formation of elongated cells at the expanding edge and was induced when cells were inoculated onto lawns of metabolically inactive Campylobacter jejuni cells, autoclaved bacterial biomass, adsorbed milk, and adsorbed blood atop hard agar plates. Phosphatidylcholine (PC), bacterial membrane components, and sterile human fecal extracts were also sufficient to induce filamentous expansion. Screening of eight other Bacillus spp. showed that filamentous motility was conserved amongst B. cereus group species to varying degrees. RNA-Seq of elongated expanding cells collected from adsorbed milk and PC lawns versus control rod-shaped cells revealed dysregulation of genes involved in metabolism and membrane transport, sporulation, quorum sensing, antibiotic synthesis, and virulence (e.g., hblA/B/C/D and plcR). These findings characterize the robustness and ecological significance of filamentous surface motility in B. cereus group species and lay the foundation for understanding the biological role it may play during environment and host colonization.

scholarly journals Topology of RbsC, the Membrane Component of the Escherichia coli Ribose Transporter

2003 ◽  
Vol 185 (17) ◽  
pp. 5234-5239 ◽  
Author(s):  
Jeffrey B. Stewart ◽  
Mark A. Hermodson
Keyword(s):  
Escherichia Coli ◽  
Membrane Protein ◽  
Fluorescence Labeling ◽  
Bacterial Cells ◽  
Membrane Component ◽  
Single Batch ◽  
Localization Information

ABSTRACT The topology of RbsC, the membrane component of the ribose transporter in Escherichia coli, has been determined by using 34 single-cysteine mutants and a modified fluorescence labeling technique designated multiplex labeling. This technique gives topology, expression, and localization information for a membrane protein from a single batch of bacterial cells. The results indicate that RbsC contains 10 transmembrane-spanning helices, with the N and C termini being in the cytosol. This topology matches predictions from the latest prediction programs and the topology of the similar, recently crystallized membrane protein BtuC.

scholarly journals Topology of AspT, the Aspartate:Alanine Antiporter of Tetragenococcus halophilus, Determined by Site-Directed Fluorescence Labeling

2007 ◽  
Vol 189 (19) ◽  
pp. 7089-7097 ◽  
Author(s):  
Kei Nanatani ◽  
Takashi Fujiki ◽  
Kazuhiko Kanou ◽  
Mayuko Takeda-Shitaka ◽  
Hideaki Umeyama ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Amino Acid Sequences ◽  
Fluorescence Labeling ◽  
Dimensional Structure ◽  
Specific Probe ◽  
Cytoplasmic Loop ◽  
Intact Cells ◽  
Tetragenococcus Halophilus ◽  
Substituted Cysteine Accessibility ◽  
And Function

ABSTRACT The gram-positive lactic acid bacterium Tetragenococcus halophilus catalyzes the decarboxylation of l-aspartate (Asp) with release of l-alanine (Ala) and CO2. The decarboxylation reaction consists of two steps: electrogenic exchange of Asp for Ala catalyzed by an aspartate:alanine antiporter (AspT) and intracellular decarboxylation of the transported Asp catalyzed by an l-aspartate-β-decarboxylase (AspD). AspT belongs to the newly classified aspartate:alanine exchanger family (transporter classification no. 2.A.81) of transporters. In this study, we were interested in the relationship between the structure and function of AspT and thus analyzed the topology by means of the substituted-cysteine accessibility method using the impermeant, fluorescent, thiol-specific probe Oregon Green 488 maleimide (OGM) and the impermeant, nonfluorescent, thiol-specific probe [2-(trimethylammonium)ethyl]methanethiosulfonate bromide. We generated 23 single-cysteine variants from a six-histidine-tagged cysteineless AspT template. A cysteine position was assigned an external location if the corresponding single-cysteine variant reacted with OGM added to intact cells, and a position was assigned an internal location if OGM labeling required cell lysis. The topology analyses revealed that AspT has a unique topology; the protein has 10 transmembrane helices (TMs), a large hydrophilic cytoplasmic loop (about 180 amino acids) between TM5 and TM6, N and C termini that face the periplasm, and a positively charged residue (arginine 76) within TM3. Moreover, the three-dimensional structure constructed by means of the full automatic modeling system indicates that the large hydrophilic cytoplasmic loop of AspT possesses a TrkA_C domain and a TrkA_C-like domain and that the three-dimensional structures of these domains are similar to each other even though their amino acid sequences show low similarity.

A study of Erythrocyte Membrane Proteins and Urinary Polypeptides in Conga Drumming Haemoglobinuria

1985 ◽  
Vol 69 (1) ◽  
pp. 105-108 ◽  
Author(s):  
Richard D. Levy ◽  
Ali A. Khaleeli ◽  
Beatrice L. Griffiths ◽  
Yvonne H. Edwards
Keyword(s):  
Membrane Proteins ◽  
Carbonic Anhydrase ◽  
Erythrocyte Membrane ◽  
Acute Phase ◽  
Protein Staining ◽  
Membrane Components ◽  
Erythrocyte Membrane Proteins ◽  
Erythrocyte Carbonic Anhydrase

1. The erythrocyte membrane proteins and glycoproteins and urinary polypeptides have been examined in a patient exhibiting intermittent pigmenturia associated with conga drumming. 2. Significant excretion of haemoglobin, albumin and probably erythrocyte carbonic anhydrase but not myoglobin occurred during the acute phase of the conga drumming-induced pigmenturia. This usually ceased within 24-48 h. 3. We found no evidence of aberrant erythrocyte membrane components on electrophoresis with either protein staining or a range of 125I-labelled lectins used for detection.

Nitric oxide induced oxidative changes in erythrocyte membrane components

2008 ◽  
Vol 32 (1) ◽  
pp. 114-120 ◽  
Author(s):  
Joanna Brzeszczynska ◽  
Krzysztof Gwozdzinski
Keyword(s):  
Nitric Oxide ◽  
Erythrocyte Membrane ◽  
Membrane Components
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