Opposite effect of membrane raft perturbation on transport activity of KCC2 and NKCC1

Anna Maria Hartmann, Peter Blaesse, Thorsten Kranz, Meike Wenz, Jens Schindler, Kai Kaila, Eckhard Friauf, Hans Gerd Nothwang

Tutkimustuotos: ArtikkelijulkaisuArtikkeliTieteellinenvertaisarvioitu

Kuvaus

In the majority of neurons, the intracellular Cl- concentration is set by the activity of the Na+-K+-2Cl- cotransporter (NKCC1) and the K+-Cl- cotransporter (KCC2). Here, we investigated the cotransporters' functional dependence on membrane rafts. In the mature rat brain, NKCC1 was mainly insoluble in Brij 58 and co-distributed with the membrane raft marker flotillin-1 in sucrose density flotation experiments. In contrast, KCC2 was found in the insoluble fraction as well as in the soluble fraction, where it co-distributed with the non-raft marker transferrin receptor. Both KCC2 populations displayed a mature glycosylation pattern. Disrupting membrane rafts with methyl-beta-cyclodextrin (M beta CD) increased the solubility of KCC2, yet had no effect on NKCC1. In human embryonic kidney-293 cells, KCC2 was strongly activated by a combined treatment with M beta CD and sphingomyelinase, while NKCC1 was inhibited. These data indicate that membrane rafts render KCC2 inactive and NKCC1 active. In agreement with this, inactive KCC2 of the perinatal rat brainstem largely partitioned into membrane rafts. In addition, the exposure of the transporters to M beta CD and sphingomyelinase showed that the two transporters differentially interact with the membrane rafts. Taken together, membrane raft association appears to represent a mechanism for co-ordinated regulation of chloride transporter function.
Alkuperäiskielienglanti
LehtiJournal of Neurochemistry
Vuosikerta111
Numero2
Sivut321-331
Sivumäärä11
ISSN0022-3042
DOI - pysyväislinkit
TilaJulkaistu - 2009
OKM-julkaisutyyppiA1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä, vertaisarvioitu

Tieteenalat

  • 311 Peruslääketieteet
  • 318 Lääketieteen bioteknologia
  • 411 Maatalous ja metsätieteet
  • 118 Biotieteet
  • 219 Ympäristön bioteknologia
  • 519 Yhteiskuntamaantiede, talousmaantiede

Lainaa tätä

Hartmann, A. M., Blaesse, P., Kranz, T., Wenz, M., Schindler, J., Kaila, K., ... Nothwang, H. G. (2009). Opposite effect of membrane raft perturbation on transport activity of KCC2 and NKCC1. Journal of Neurochemistry, 111(2), 321-331. https://doi.org/10.1111/j.1471-4159.2009.06343.x
Hartmann, Anna Maria ; Blaesse, Peter ; Kranz, Thorsten ; Wenz, Meike ; Schindler, Jens ; Kaila, Kai ; Friauf, Eckhard ; Nothwang, Hans Gerd. / Opposite effect of membrane raft perturbation on transport activity of KCC2 and NKCC1. Julkaisussa: Journal of Neurochemistry. 2009 ; Vuosikerta 111, Nro 2. Sivut 321-331.
@article{d4988d9fbff24667b473502d0c859e34,
title = "Opposite effect of membrane raft perturbation on transport activity of KCC2 and NKCC1",
abstract = "In the majority of neurons, the intracellular Cl- concentration is set by the activity of the Na+-K+-2Cl- cotransporter (NKCC1) and the K+-Cl- cotransporter (KCC2). Here, we investigated the cotransporters' functional dependence on membrane rafts. In the mature rat brain, NKCC1 was mainly insoluble in Brij 58 and co-distributed with the membrane raft marker flotillin-1 in sucrose density flotation experiments. In contrast, KCC2 was found in the insoluble fraction as well as in the soluble fraction, where it co-distributed with the non-raft marker transferrin receptor. Both KCC2 populations displayed a mature glycosylation pattern. Disrupting membrane rafts with methyl-beta-cyclodextrin (M beta CD) increased the solubility of KCC2, yet had no effect on NKCC1. In human embryonic kidney-293 cells, KCC2 was strongly activated by a combined treatment with M beta CD and sphingomyelinase, while NKCC1 was inhibited. These data indicate that membrane rafts render KCC2 inactive and NKCC1 active. In agreement with this, inactive KCC2 of the perinatal rat brainstem largely partitioned into membrane rafts. In addition, the exposure of the transporters to M beta CD and sphingomyelinase showed that the two transporters differentially interact with the membrane rafts. Taken together, membrane raft association appears to represent a mechanism for co-ordinated regulation of chloride transporter function.",
keywords = "311 Basic medicine, 318 Medical biotechnology, 411 Agriculture and forestry, 118 Biological sciences, 219 Environmental biotechnology, 519 Social and economic geography",
author = "Hartmann, {Anna Maria} and Peter Blaesse and Thorsten Kranz and Meike Wenz and Jens Schindler and Kai Kaila and Eckhard Friauf and Nothwang, {Hans Gerd}",
year = "2009",
doi = "10.1111/j.1471-4159.2009.06343.x",
language = "English",
volume = "111",
pages = "321--331",
journal = "Journal of Neurochemistry",
issn = "0022-3042",
publisher = "LIPPINCOTT-RAVEN PUBL",
number = "2",

}

Hartmann, AM, Blaesse, P, Kranz, T, Wenz, M, Schindler, J, Kaila, K, Friauf, E & Nothwang, HG 2009, 'Opposite effect of membrane raft perturbation on transport activity of KCC2 and NKCC1', Journal of Neurochemistry, Vuosikerta 111, Nro 2, Sivut 321-331. https://doi.org/10.1111/j.1471-4159.2009.06343.x

Opposite effect of membrane raft perturbation on transport activity of KCC2 and NKCC1. / Hartmann, Anna Maria; Blaesse, Peter; Kranz, Thorsten; Wenz, Meike; Schindler, Jens; Kaila, Kai; Friauf, Eckhard; Nothwang, Hans Gerd.

julkaisussa: Journal of Neurochemistry, Vuosikerta 111, Nro 2, 2009, s. 321-331.

Tutkimustuotos: ArtikkelijulkaisuArtikkeliTieteellinenvertaisarvioitu

TY - JOUR

T1 - Opposite effect of membrane raft perturbation on transport activity of KCC2 and NKCC1

AU - Hartmann, Anna Maria

AU - Blaesse, Peter

AU - Kranz, Thorsten

AU - Wenz, Meike

AU - Schindler, Jens

AU - Kaila, Kai

AU - Friauf, Eckhard

AU - Nothwang, Hans Gerd

PY - 2009

Y1 - 2009

N2 - In the majority of neurons, the intracellular Cl- concentration is set by the activity of the Na+-K+-2Cl- cotransporter (NKCC1) and the K+-Cl- cotransporter (KCC2). Here, we investigated the cotransporters' functional dependence on membrane rafts. In the mature rat brain, NKCC1 was mainly insoluble in Brij 58 and co-distributed with the membrane raft marker flotillin-1 in sucrose density flotation experiments. In contrast, KCC2 was found in the insoluble fraction as well as in the soluble fraction, where it co-distributed with the non-raft marker transferrin receptor. Both KCC2 populations displayed a mature glycosylation pattern. Disrupting membrane rafts with methyl-beta-cyclodextrin (M beta CD) increased the solubility of KCC2, yet had no effect on NKCC1. In human embryonic kidney-293 cells, KCC2 was strongly activated by a combined treatment with M beta CD and sphingomyelinase, while NKCC1 was inhibited. These data indicate that membrane rafts render KCC2 inactive and NKCC1 active. In agreement with this, inactive KCC2 of the perinatal rat brainstem largely partitioned into membrane rafts. In addition, the exposure of the transporters to M beta CD and sphingomyelinase showed that the two transporters differentially interact with the membrane rafts. Taken together, membrane raft association appears to represent a mechanism for co-ordinated regulation of chloride transporter function.

AB - In the majority of neurons, the intracellular Cl- concentration is set by the activity of the Na+-K+-2Cl- cotransporter (NKCC1) and the K+-Cl- cotransporter (KCC2). Here, we investigated the cotransporters' functional dependence on membrane rafts. In the mature rat brain, NKCC1 was mainly insoluble in Brij 58 and co-distributed with the membrane raft marker flotillin-1 in sucrose density flotation experiments. In contrast, KCC2 was found in the insoluble fraction as well as in the soluble fraction, where it co-distributed with the non-raft marker transferrin receptor. Both KCC2 populations displayed a mature glycosylation pattern. Disrupting membrane rafts with methyl-beta-cyclodextrin (M beta CD) increased the solubility of KCC2, yet had no effect on NKCC1. In human embryonic kidney-293 cells, KCC2 was strongly activated by a combined treatment with M beta CD and sphingomyelinase, while NKCC1 was inhibited. These data indicate that membrane rafts render KCC2 inactive and NKCC1 active. In agreement with this, inactive KCC2 of the perinatal rat brainstem largely partitioned into membrane rafts. In addition, the exposure of the transporters to M beta CD and sphingomyelinase showed that the two transporters differentially interact with the membrane rafts. Taken together, membrane raft association appears to represent a mechanism for co-ordinated regulation of chloride transporter function.

KW - 311 Basic medicine

KW - 318 Medical biotechnology

KW - 411 Agriculture and forestry

KW - 118 Biological sciences

KW - 219 Environmental biotechnology

KW - 519 Social and economic geography

U2 - 10.1111/j.1471-4159.2009.06343.x

DO - 10.1111/j.1471-4159.2009.06343.x

M3 - Article

VL - 111

SP - 321

EP - 331

JO - Journal of Neurochemistry

JF - Journal of Neurochemistry

SN - 0022-3042

IS - 2

ER -