Highly Sensitive and Stable Phosphatidylserine Liposome Aggregation Assay for Annexins

Lee, George; Pollard, Harvey B.

doi:10.1006/abio.1997.2311

Cited by 24 publications

(15 citation statements)

References 28 publications

(3 reference statements)

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“…In the presence of membranes Annexin-V self-assembles into a trimers that form a 2-dimensional crystal lattice. Consistent with these structural data, Annexin-I aggregates PtdSer vesicles or chromaffin granules, whereas Annexin-V does not (149, 150). This also explains why Annexin-V plays an anti-coagulant role in the blood: the 2-D crystal of Annexin-V essentially masks PtdSer that may be externalized at low levels on cells.…”

Section: Introductionsupporting

confidence: 73%

Anoctamins/TMEM16 Proteins: Chloride Channels Flirting with Lipids and Extracellular Vesicles

Whitlock

Hartzell

2017

Annu. Rev. Physiol.

147

159

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Anoctamin/TMEM16 proteins exhibit diverse functions in cells throughout the body and are implicated in several human diseases. Although the founding members ANO1 (TMEM16A) and ANO2 (TMEM16B) are Ca2+-activated Cl− channels, most ANO paralogs are Ca2+-dependent phospholipid scramblases that serve as channels that facilitate the movement (“scrambling”) of phospholipids between leaflets of the membrane bilayer. Phospholipid scrambling significantly alters the physical properties of the membrane and its landscape and has vast downstream signaling consequences. In particular, phosphatidylserine exposed on the external leaflet of the plasma membrane functions as a ligand for receptors vital for cell-cell communication. A major consequence of Ca2+-dependent scrambling is the release of extracellular vesicles that function as intercellular messengers by delivering signaling proteins and non-coding RNAs to alter target cell function. We discuss the physiological implications of Ca2+-dependent phospholipid scrambling, the extracellular vesicles associated with this activity, and the roles of ANOs in these processes.

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Section: Introductionsupporting

confidence: 73%

Anoctamins/TMEM16 Proteins: Chloride Channels Flirting with Lipids and Extracellular Vesicles

Whitlock

Hartzell

2017

Annu. Rev. Physiol.

147

159

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“…Aggregation of unsaturated POPS and DOPG liposomes, induced by interaction of the hydrophobic hexane-dibutyl structure of 3 with the unsaturated lipid bilayer, resulting in enhanced binding and catalysis by the larger phospholipid aggregates formed, may be responsible, at least in part, for the kinetic behavior displayed. Aggregation and fusion of phosphatidylserine and phosphatidylglycerol liposomes, induced by a variety of reagents, has been previously reported [27][28][29].…”

Section: Discussionmentioning

confidence: 92%

Diazeniumdiolate reactivity in model membrane systems

et al. 2008

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The effect of small unilamellar phospholipid vesicles on the acid-catalyzed dissociation of nitric oxide from diazeniumdiolate ions, R 1 R 2 N[N(O)NO] -, [1: R 1 = H 2 N(CH 2 ) 3 -, R 2 = H 2 N(CH 2 ) 3 NH (CH 2 ) 4 -; 2: R 1 = R 2 = H 2 N(CH 2 ) 3 -; 3: R 1 = n-butyl-, R 2 = n-butyl-NH 2 + (CH 2 ) 6 -; 4: R 1 = R 2 = n Pr-] has been examined at pH 7.4 and 37 °C. NO release was catalyzed by anionic liposomes (DPPG, DOPG, DMPS, POPS and DOPA) and by mixed phosphatidylglycerol/phosphatidylcholine (DPPG/ DPPC and DOPG/DPPC) covesicles, while cationic liposomes derived from 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) and the zwitterionic liposome DMPC did not significantly affect the dissociation rates of the substrates examined. Enhancement of the dissociation rate constant in DPPG liposome media (0.010M phosphate buffer, pH 7.4, 37 °C) at 10 mM phosphoglycerol levels, ranged from 37 for 1 to 1.2 for the anionic diazeniumdiolate 4, while DOPA effected the greatest rate enhancement, achieving 49-fold rate increases with 1 under similar conditions. The observed catalysis decreases with increase in the bulk concentration of electrolytes in the reaction media. Quantitative analysis of catalytic effects has been obtained through the application of pseudophase kinetic models and equilibrium binding constants at different liposome interfaces are compared. The stoichiometry of nitric oxide release from 1 and 2 in DPPG/DPPC liposome media has been obtained through oxyhemoglobin assay. DPPG = 1,2

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“…Although a prior study suggested that WNV induced cell death in the N2a neuroblastoma cell line (48) through an apoptotic mechanism, it was unclear whether this occurred in primary neurons. To evaluate this, ESNC were infected and analyzed kinetically for the earliest stages of apoptosis by annexin V staining (36,37). Because annexin V also binds to phosphatidylserine on the inner leaflet of the membrane of necrotic cells that have lost membrane integrity, cells were counterstained with 7-AAD to distinguish viable apoptotic cells from dead cells by two-color flow cytometry.…”

Section: Wnv Infection In Esncmentioning

confidence: 99%

Infection and Injury of Neurons by West NileEncephalitisVirus

Shrestha¹,

Gottlieb

Diamond

2003

J Virol

219

173

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West Nile virus (WNV) infects neurons and leads to encephalitis, paralysis, and death in humans, animals, and birds. We investigated the mechanism by which neuronal injury occurs after WNV infection. Neurons in the anterior horn of the spinal cords of paralyzed mice exhibited a high degree of WNV infection, leukocyte infiltration, and degeneration. Because it was difficult to distinguish whether neuronal injury was caused by viral infection or by the immune system response, a novel tissue culture model for WNV infection was established in neurons derived from embryonic stem (ES) cells. Undifferentiated ES cells were relatively resistant to WNV infection. After differentiation, ES cells expressed neural antigens, acquired a neuronal phenotype, and became permissive for WNV infection. Within 48 h of exposure to an exceedingly low multiplicity of infection (5 ؋ 10 ؊4 ), 50% of ES cell-derived neurons became infected, producing nearly 10 7 PFU of infectious virus per ml, and began to die by an apoptotic mechanism. The establishment of a tractable virus infection model in ES cell-derived neurons facilitates the study of the molecular basis of neurotropism and the mechanisms of viral and immune-mediated neuronal injury after infection by WNV or other neurotropic pathogens.West Nile virus (WNV) is a neurotropic flavivirus that is transmitted by mosquitoes and causes West Nile encephalitis in humans, animals, and birds (35). Humans develop a febrile illness, with a subset of cases progressing to meningoencephalitis (56) or a paralytic or polio-like syndrome (38; T. J. John, Letter, N. Engl. J. Med. 348:564-566, 2003). WNV causes paralysis (14, 71), in part by destroying neurons in the anterior horn of the spinal cord, where motor neurons reside (19,38). Although neuronal injury may be directly caused by viral infection (11, 71), destruction has also been attributed to infiltrating leukocytes, inflammatory cytokines, and activated microglial cells (19,20,24,61).In principle, tissue culture models of viral infection in primary neurons can distinguish injury that is caused by virus from injury that is caused by the immune response. For many neurotropic viruses (e.g., poliovirus, herpes simplex virus type 1, Japanese encephalitis virus, rabies virus, and Sindbis virus), cells from neuroblastomas and primary cultures from embryonic or neonatal mice and rats have been used as models of neuronal infection (11,22,28,34,40,63). However, the existing primary culture systems have limitations, as they are difficult to establish and scale up for high-throughput applications. In addition, the cultures often contain cells of multiple neuronal cell types, and genetic manipulation is constrained in these postmitotic cells.Embryonic stem (ES) cells are totipotent continuous cell lines that can be differentiated into neural, muscle, and hematopoietic cells (1, 27, 68, 70) and manipulated genetically (10, 50). We and others have efficiently differentiated ES cells into neurons (ESNC) after retinoic acid induction or by lineage selection (1...

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Highly Sensitive and Stable Phosphatidylserine Liposome Aggregation Assay for Annexins

Cited by 24 publications

References 28 publications

Anoctamins/TMEM16 Proteins: Chloride Channels Flirting with Lipids and Extracellular Vesicles

Anoctamins/TMEM16 Proteins: Chloride Channels Flirting with Lipids and Extracellular Vesicles

Diazeniumdiolate reactivity in model membrane systems

Infection and Injury of Neurons by West NileEncephalitisVirus

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