Artículo

Leal Denis, M.F.; Lefevre, S.D.; Alvarez, C.L.; Lauri, N.; Enrique, N.; Rinaldi, D.E.; Gonzalez-Lebrero, R.; Vecchio, L.E.; Espelt, M.V.; Stringa, P.; Muñoz-Garay, C.; Milesi, V.; Ostuni, M.A.; Herlax, V.; Schwarzbaum, P.J. "Regulation of extracellular ATP of human erythrocytes treated with α-hemolysin. Effects of cell volume, morphology, rheology and hemolysis" (2019) Biochimica et Biophysica Acta - Molecular Cell Research. 1866(5):896-915
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Abstract:

Alpha-hemolysin (HlyA) of uropathogenic strains of Escherichia coli irreversibly binds to human erythrocytes (RBCs) and triggers activation of ATP release and metabolic changes ultimately leading to hemolysis. We studied the regulation of extracellular ATP (ATPe) of RBCs exposed to HlyA. Luminometry was used to assess ATP release and ATPe hydrolysis, whereas changes in cell volume and morphology were determined by electrical impedance, ektacytometry and aggregometry. Exposure of RBCs to HlyA induced a strong increase of [ATPe] (3–36-fold) and hemolysis (1–44-fold), partially compensated by [ATPe] hydrolysis by ectoATPases and intracellular ATPases released by dead cells. Carbenoxolone, a pannexin 1 inhibitor, partially inhibited ATP release (43–67%). The un-acylated toxin ProHlyA and the deletion analog HlyA∆914-936 were unable to induce ATP release or hemolysis. For HlyA treated RBCs, a data driven mathematical model showed that simultaneous lytic and non-lytic release mainly governed ATPe kinetics, while ATPe hydrolysis became important after prolonged toxin exposure. HlyA induced a 1.5-fold swelling, while blocking this swelling reduced ATP release by 77%. Blocking ATPe activation of purinergic P2X receptors reduced swelling by 60–80%. HlyA-RBCs showed an acute 1.3–2.2-fold increase of Ca 2+ i, increased crenation and externalization of phosphatidylserine. Perfusion of HlyA-RBCs through adhesion platforms showed strong adhesion to activated HMEC cells, followed by rapid detachment. HlyA exposed RBCs exhibited increased sphericity under osmotic stress, reduced elongation under shear stress, and very low aggregation in viscous media. Overall results showed that HlyA-RBCs displayed activated ATP release, high but weak adhesivity, low deformability and aggregability and high sphericity. © 2019 Elsevier B.V.

Registro:

Documento: Artículo
Título:Regulation of extracellular ATP of human erythrocytes treated with α-hemolysin. Effects of cell volume, morphology, rheology and hemolysis
Autor:Leal Denis, M.F.; Lefevre, S.D.; Alvarez, C.L.; Lauri, N.; Enrique, N.; Rinaldi, D.E.; Gonzalez-Lebrero, R.; Vecchio, L.E.; Espelt, M.V.; Stringa, P.; Muñoz-Garay, C.; Milesi, V.; Ostuni, M.A.; Herlax, V.; Schwarzbaum, P.J.
Filiación:Universidad de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas, Instituto de Química y Fisico-Química Biológicas (IQUIFIB) “Prof. Alejandro C. Paladini”, Facultad de Farmacia y Bioquímica, Buenos Aires, Junín 956, Argentina
Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Departamento de Química Analítica, Cátedra de Química Química Analítica y Fisicoquímica, Buenos Aires, Junín 956, Argentina
UMR-S1134, Integrated Biology of Red Blood Cells, INSERM, Université Paris Diderot, Sorbonne Paris Cité, Université de la Réunion, Université des Antilles, Paris, F-75015, France
Institut National de la Transfusion Sanguine, Laboratoire d'Excellence GR-Ex, Paris, F-75015, France
Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Biodiversidad y Biología Experimental, Buenos Aires, Intendente Güiraldes 2160, Argentina
Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Química Biológica. Cátedra de Química Biológica Superior, Buenos Aires, Junín 956, Argentina
Universidad Nacional de La Plata, Consejo Nacional de Investigaciones Científicas y Técnicas, Instituto de Estudios Inmunológicos y Fisiopatológicos (IIFP), Facultad de Ciencias Exactas, La Plata, Calle 47 y 115, Argentina
Universidad Nacional de la Plata, Facultad de Ciencias Exactas, Departamento de Ciencias Biológicas, Cátedra de Fisiología, Calle 47, Casco Urbano, La Plata, Argentina
Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Departamento de Química Biológica, Cátedra de Química Biológica, Buenos Aires, Junín 956, Argentina
Universidad Favaloro, Consejo Nacional de Investigaciones Científicas y Técnicas, Instituto de Medicina Traslacional, Trasplante y Bioingeniería (IMETTyB), Av. Entre Ríos 495, Buenos Aires, Argentina
Universidad Nacional de La Plata, Laboratorio de Trasplante de Órganos y Tejidos, Facultad de Ciencias, Calle 60 y 120, La Plata, Argentina
Instituto de Ciencias Físicas, Universidad Nacional Autónoma de México (UNAM), Av. Universidad s/n, Cuernavaca, Mexico
Universidad Nacional de La Plata, Consejo Nacional de Investigaciones Científicas y Técnicas, Instituto de Investigaciones Bioquímicas de La Plata (INIBIOLP) “Prof. Dr. Rodolfo R. Brenner”, Facultad de Ciencias Médicas, Av. 60 y Av. 120, La Plata, Argentina
Universidad Nacional de La Plata, Facultad de Ciencias Médicas, Av. 60 y Av. 120, La Plata, Argentina
Palabras clave:Adhesion, aggregation; ATP-transport; E.coli; hemolysin; Hemolysis; adenosine triphosphatase; alpha hemolysin; apyrase; calcium; carbenoxolone; hemoglobin; liposome; purinergic P2X receptor; animal experiment; Article; calcium cell level; cell aggregation; cell isolation; cell shape; cell structure; cell swelling; cell volume; erythrocyte; flow kinetics; hemolysis; hemolysis assay; human; human cell; hydrolysis; impedance; male; mathematical model; membrane potential; mesenteric arterial bed; nonhuman; osmotic stress; priority journal; protein purification; protein secretion; rat; refraction index; regulatory mechanism; scanning electron microscopy; shear stress
Año:2019
Volumen:1866
Número:5
Página de inicio:896
Página de fin:915
DOI: http://dx.doi.org/10.1016/j.bbamcr.2019.01.018
Título revista:Biochimica et Biophysica Acta - Molecular Cell Research
Título revista abreviado:Biochim. Biophys. Acta Mol. Cell Res.
ISSN:01674889
CODEN:BAMRD
CAS:adenosine triphosphatase, 37289-25-1, 9000-83-3; apyrase, 9000-95-7; calcium, 7440-70-2, 14092-94-5; carbenoxolone, 5697-56-3, 7421-40-1; hemoglobin, 9008-02-0
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01674889_v1866_n5_p896_LealDenis

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Citas:

---------- APA ----------
Leal Denis, M.F., Lefevre, S.D., Alvarez, C.L., Lauri, N., Enrique, N., Rinaldi, D.E., Gonzalez-Lebrero, R.,..., Schwarzbaum, P.J. (2019) . Regulation of extracellular ATP of human erythrocytes treated with α-hemolysin. Effects of cell volume, morphology, rheology and hemolysis. Biochimica et Biophysica Acta - Molecular Cell Research, 1866(5), 896-915.
http://dx.doi.org/10.1016/j.bbamcr.2019.01.018
---------- CHICAGO ----------
Leal Denis, M.F., Lefevre, S.D., Alvarez, C.L., Lauri, N., Enrique, N., Rinaldi, D.E., et al. "Regulation of extracellular ATP of human erythrocytes treated with α-hemolysin. Effects of cell volume, morphology, rheology and hemolysis" . Biochimica et Biophysica Acta - Molecular Cell Research 1866, no. 5 (2019) : 896-915.
http://dx.doi.org/10.1016/j.bbamcr.2019.01.018
---------- MLA ----------
Leal Denis, M.F., Lefevre, S.D., Alvarez, C.L., Lauri, N., Enrique, N., Rinaldi, D.E., et al. "Regulation of extracellular ATP of human erythrocytes treated with α-hemolysin. Effects of cell volume, morphology, rheology and hemolysis" . Biochimica et Biophysica Acta - Molecular Cell Research, vol. 1866, no. 5, 2019, pp. 896-915.
http://dx.doi.org/10.1016/j.bbamcr.2019.01.018
---------- VANCOUVER ----------
Leal Denis, M.F., Lefevre, S.D., Alvarez, C.L., Lauri, N., Enrique, N., Rinaldi, D.E., et al. Regulation of extracellular ATP of human erythrocytes treated with α-hemolysin. Effects of cell volume, morphology, rheology and hemolysis. Biochim. Biophys. Acta Mol. Cell Res. 2019;1866(5):896-915.
http://dx.doi.org/10.1016/j.bbamcr.2019.01.018