scholarly journals Characterization of Microvesicles Released from Human Red Blood Cells

2016 ◽  
Vol 38 (3) ◽  
pp. 1085-1099 ◽  
Author(s):  
Duc Bach Nguyen ◽  
Thi Bich Thuy Ly ◽  
Mauro Carlos Wesseling ◽  
Marius Hittinger ◽  
Afra Torge ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Zeta Potential ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Annexin V ◽  
Human Red Blood Cells ◽  
Isolation And Characterization ◽  
Kinase C

Background/Aims: Extracellular vesicles (EVs) are spherical fragments of cell membrane released from various cell types under physiological as well as pathological conditions. Based on their size and origin, EVs are classified as exosome, microvesicles (MVs) and apoptotic bodies. Recently, the release of MVs from human red blood cells (RBCs) under different conditions has been reported. MVs are released by outward budding and fission of the plasma membrane. However, the outward budding process itself, the release of MVs and the physical properties of these MVs have not been well investigated. The aim of this study is to investigate the formation process, isolation and characterization of MVs released from RBCs under conditions of stimulating Ca2+ uptake and activation of protein kinase C. Methods: Experiments were performed based on single cell fluorescence imaging, fluorescence activated cell sorter/flow cytometer (FACS), scanning electron microscopy (SEM), atomic force microscopy (AFM) and dynamic light scattering (DLS). The released MVs were collected by differential centrifugation and characterized in both their size and zeta potential. Results: Treatment of RBCs with 4-bromo-A23187 (positive control), lysophosphatidic acid (LPA), or phorbol-12 myristate-13 acetate (PMA) in the presence of 2 mM extracellular Ca2+ led to an alteration of cell volume and cell morphology. In stimulated RBCs, exposure of phosphatidylserine (PS) and formation of MVs were observed by using annexin V-FITC. The shedding of MVs was also observed in the case of PMA treatment in the absence of Ca2+, especially under the transmitted bright field illumination. By using SEM, AFM and DLS the morphology and size of stimulated RBCs, MVs were characterized. The sizes of the two populations of MVs were 205.8 ± 51.4 nm and 125.6 ± 31.4 nm, respectively. Adhesion of stimulated RBCs and MVs was observed. The zeta potential of MVs was determined in the range from - 40 mV to - 10 mV depended on the solutions and buffers used. Conclusion: An increase of intracellular Ca2+ or an activation of protein kinase C leads to the formation and release of MVs in human RBCs.

scholarly journals Supernatants and lipids from stored red blood cells activate pulmonary microvascular endothelium through the BLT2 receptor and protein kinase C activation

Transfusion ◽  
2017 ◽  
Vol 57 (11) ◽  
pp. 2690-2700 ◽  
Author(s):  
Christopher C. Silliman ◽  
Marguerite R. Kelher ◽  
Samina Y. Khan ◽  
F. Bernadette West ◽  
Nathan J.D. McLaughlin ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Microvascular Endothelium ◽  
Kinase C ◽  
Protein Kinase C Activation

Isolation and characterization of PKC-L, a new member of the protein kinase C-related gene family specifically expressed in lung, skin, and heart

1991 ◽  
Vol 11 (1) ◽  
pp. 126-133 ◽  
Author(s):  
N Bacher ◽  
Y Zisman ◽  
E Berent ◽  
E Livneh
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Gene Family ◽  
Phorbol Esters ◽  
Structural Features ◽  
Cos Cells ◽  
Isolation And Characterization ◽  
Kinase C ◽  
Human Epidermoid Carcinoma

We have isolated and characterized a new human cDNA, coding for a protein kinase, related to the protein kinase C (PKC) gene family. Although this protein kinase shares some homologous sequences and structural features with the four members of the PKC family initially isolated (alpha, beta I, beta II, and gamma), it shows more homology with the recently described PKC-related subfamily, encoded by the cDNAs delta, epsilon, and zeta. The transcript for this gene product, termed PKC-L, is most abundant in lung tissue, less expressed in heart and skin tissue, and exhibited very low expression in brain tissue. Thus, its tissue distribution is different from that described for other mammalian members of the PKC gene family, their expression being enriched in brain tissues. PKC-L is also expressed in several human cell lines, including the human epidermoid carcinoma line A431. The ability of phorbol esters to bind to and stimulate the kinase activity of PKC-L was revealed by introducing the cDNA into COS cells.

Isolation and characterization of two new Drosophila protein kinase C genes, including one specifically expressed in photoreceptor cells

Cell ◽  
1989 ◽  
Vol 57 (3) ◽  
pp. 403-412 ◽  
Author(s):  
Eric Schaeffer ◽  
Dean Smith ◽  
Graeme Mardon ◽  
William Quinn ◽  
Charles Zuker
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Photoreceptor Cells ◽  
Isolation And Characterization ◽  
Kinase C ◽  
Drosophila Protein

scholarly journals Isolation and characterization of an age-related antigen present on senescent human red blood cells

Blood ◽  
1981 ◽  
Vol 58 (2) ◽  
pp. 341-349
Author(s):  
EM Alderman ◽  
HH Fudenberg ◽  
RE Lovins
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Double Immunodiffusion ◽  
Specific Density ◽  
Human Red Blood Cells ◽  
Rbc Membrane ◽  
Isolation And Characterization ◽  
Age Related ◽  
Membrane Bound

Autologous membrane-bound IgG was isolated from a subpopulation of human red blood cells (RBC) with specific density greater than 1.110, by affinity chromatography of purified RBC membrane glycoprotein preparations using immobilized wheat germ agglutinin and immobilized anti-human immunoglobulin (Ig) as immunoabsorbents. The Ig-containing population thus obtained, when further separated by chromatography on Sephadex G-200 in the presence of chaotropic agents, yielded four peaks (Ia, Ib, II, and III). Double immunodiffusion revealed the presence of Ig in the first three peaks (IgM in peak Ia, IgA in Ib, and IgG in II) but not in peak III. Peak III was precipitated by the Ig-containing peaks (Ia, Ib, and II) in immunodiffusion assays, suggesting that the antigenic membrane determinants responsible for the binding of autologous Ig to senescent human RBC were contained in this peak (III). Peaks Ia, Ib and II precipitate purified asialoglycophorin; peak III was reactive with purified autoantibodies directed against asialoglycophorin. These results suggest that an age-related antigenic determinant(s) present on senescent human RBC is exposed by desialylation of the major sialoglycoprotein component of the RBC membrane.

Isolation and characterization of protein kinase C from human placenta

Placenta ◽  
1990 ◽  
Vol 11 (1) ◽  
pp. 27-33 ◽  
Author(s):  
C. Tertrin-Clary ◽  
M.C. Chenut ◽  
P. De la Llosa
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Human Placenta ◽  
Isolation And Characterization ◽  
Kinase C

Phorbol ester stimulates a protein kinase C–mediated agatoxin-TK–sensitive calcium permeability pathway in human red blood cells

Blood ◽  
2002 ◽  
Vol 100 (9) ◽  
pp. 3392-3399 ◽  
Author(s):  
Dina A. Andrews ◽  
Lu Yang ◽  
Philip S. Low
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Calcium Channel ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Mature Erythrocyte ◽  
Phospholipid Scramblase ◽  
Rbc Membrane ◽  
Kinase C ◽  
Calcium Permeability

AbstractCalcium entry into mature erythrocytes (red blood cells; RBCs) is associated with multiple changes in cell properties. At low intracellular Ca2+, efflux of potassium and water predominates, leading to changes in erythrocyte rheology. At higher Ca2+ content, activation of kinases and phosphatases, rupture of membrane-to-skeleton bridges, stimulation of a phospholipid scramblase and phospholipase C, and induction of transglutaminase-mediated protein cross-linking are also observed. Because the physiologic relevance of these latter responses depends partially on whether Ca2+ entry involves a regulated channel or nonspecific leak, we explored mechanisms that initiate controlled Ca2+ influx. Protein kinase C (PKC) was considered a prime candidate for the pathway regulator, and phorbol-12 myristate-13 acetate (PMA), a stimulator of PKC, was examined for its influence on erythrocyte Ca2+. PMA was found to stimulate a rapid, dose-dependent influx of calcium, as demonstrated by the increased fluorescence of an entrapped Ca2+-sensitive dye, Fluo-3/am. The PMA-induced entry was inhibited by staurosporine and the PKC-selective inhibitor chelerythrine chloride, but was activated by the phosphatase inhibitors okadaic acid and calyculin A. The PMA-promoted calcium influx was also inhibited by ω-agatoxin-TK, a calcium channel blocker specific for Cav2.1 channels. To confirm that a Cav2.1-like calcium channel exists in the mature erythrocyte membrane, RBC membrane preparations were immunoblotted with antiserum against the α1A subunit of the channel. A polypeptide of the expected molecular weight (190 kDa) was visualized. These studies indicate that an ω-agatoxin-TK–sensitive, Cav2.1-like calcium permeability pathway is present in the RBC membrane and that it may function under the control of kinases and phosphatases.

scholarly journals Isolation and characterization of frog retina protein kinase C

1990 ◽  
Vol 52 ◽  
pp. 364
Author(s):  
Motomu Terasawa ◽  
Masatoshi Hagiwara ◽  
Takahisa Hachiya ◽  
Ryoji Kobayashi ◽  
Hiroyoshi Hidaka
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Isolation And Characterization ◽  
Kinase C ◽  
Frog Retina

Protein Kinase C Activation Induces Phosphatidylserine Exposure on Red Blood Cells†

Biochemistry ◽  
2002 ◽  
Vol 41 (41) ◽  
pp. 12562-12567 ◽  
Author(s):  
Kitty de Jong ◽  
Michael P. Rettig ◽  
Philip S. Low ◽  
Frans A. Kuypers
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Kinase C ◽  
Phosphatidylserine Exposure ◽  
Protein Kinase C Activation
Sign in / Sign up

Export Citation Format

Share Document