Reassembly of the tight junction after oxidative stress depends on tyrosine kinase activity
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Reassembly of the tight junction after oxidative stress depends on tyrosine kinase activity. / Meyer, T N; Meyer-Schwesinger, Catherine; Ye, J; Denker, B M; Nigam, S K.
in: J BIOL CHEM, Jahrgang 276, Nr. 25, 22.06.2001, S. 22048-55.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Reassembly of the tight junction after oxidative stress depends on tyrosine kinase activity
AU - Meyer, T N
AU - Meyer-Schwesinger, Catherine
AU - Ye, J
AU - Denker, B M
AU - Nigam, S K
PY - 2001/6/22
Y1 - 2001/6/22
N2 - Oxidative stress compromises the tight junction, but the mechanisms underlying its recovery remain unclear. We developed a model in which oxidative stress reversibly disrupts the tight junction. Exposure of Madin-Darby canine kidney cells to hydrogen peroxide markedly reduced transepithelial resistance and disrupted the staining patterns of the tight junction proteins ZO-1 and occludin. These changes were reversed by catalase. The short-term reassembly of tight junctions was not dependent on new protein synthesis, suggesting that recovery occurs through re-utilization of existing proteins. Although ATP levels were reduced, the reduction was insufficient to explain the observed changes, since a comparable reduction of ATP levels (with 2-deoxy-D-glucose) did not induce these changes. The intracellular hydrogen peroxide scavenger pyruvate protected Madin-Darby canine kidney cells from loss of transepithelial resistance as did the heavy metal scavenger N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine. Of a wide variety of agents examined, only tyrosine kinase inhibitors and protein kinase C inhibitors markedly inhibited tight junction reassembly. During reassembly, tyrosine phosphorylation in or near the lateral membrane, was detected by immunofluorescence. The tyrosine kinase inhibitors genistein and PP-2 inhibited the recovery of transepithelial resistance and perturbed the relocalization of ZO-1 and occludin to the tight junction, indicating that tyrosine kinases, possibly members of the Src family, are critical for reassembly after oxidative stress.
AB - Oxidative stress compromises the tight junction, but the mechanisms underlying its recovery remain unclear. We developed a model in which oxidative stress reversibly disrupts the tight junction. Exposure of Madin-Darby canine kidney cells to hydrogen peroxide markedly reduced transepithelial resistance and disrupted the staining patterns of the tight junction proteins ZO-1 and occludin. These changes were reversed by catalase. The short-term reassembly of tight junctions was not dependent on new protein synthesis, suggesting that recovery occurs through re-utilization of existing proteins. Although ATP levels were reduced, the reduction was insufficient to explain the observed changes, since a comparable reduction of ATP levels (with 2-deoxy-D-glucose) did not induce these changes. The intracellular hydrogen peroxide scavenger pyruvate protected Madin-Darby canine kidney cells from loss of transepithelial resistance as did the heavy metal scavenger N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine. Of a wide variety of agents examined, only tyrosine kinase inhibitors and protein kinase C inhibitors markedly inhibited tight junction reassembly. During reassembly, tyrosine phosphorylation in or near the lateral membrane, was detected by immunofluorescence. The tyrosine kinase inhibitors genistein and PP-2 inhibited the recovery of transepithelial resistance and perturbed the relocalization of ZO-1 and occludin to the tight junction, indicating that tyrosine kinases, possibly members of the Src family, are critical for reassembly after oxidative stress.
KW - Adenosine Triphosphate
KW - Adherens Junctions
KW - Animals
KW - Cell Line
KW - Chelating Agents
KW - Connexins
KW - Dogs
KW - Genistein
KW - Hydrogen Peroxide
KW - Immunohistochemistry
KW - Membrane Potentials
KW - Metals, Heavy
KW - Oxidative Stress
KW - Protein-Tyrosine Kinases
U2 - 10.1074/jbc.M011477200
DO - 10.1074/jbc.M011477200
M3 - SCORING: Journal article
C2 - 11294856
VL - 276
SP - 22048
EP - 22055
JO - J BIOL CHEM
JF - J BIOL CHEM
SN - 0021-9258
IS - 25
ER -