Lysophosphatidic acid induces exocytic trafficking of Na+/H + exchanger 3 by E3KARP-dependent activation of phospholipase C

Jung Woong Choi; Whaseon Lee-Kwon; Eun Su Jeon; Yong Jung Kang; Kazuya Kawano; Hyeon Soo Kim; Pann Ghill Suh; Mark Donowitz; Jae Ho Kim

doi:10.1016/j.bbalip.2004.04.005

Lysophosphatidic acid induces exocytic trafficking of Na⁺/H ⁺ exchanger 3 by E3KARP-dependent activation of phospholipase C

Jung Woong Choi, Whaseon Lee-Kwon, Eun Su Jeon, Yong Jung Kang, Kazuya Kawano, Hyeon Soo Kim, Pann Ghill Suh, Mark Donowitz, Jae Ho Kim

Research output: Contribution to journal › Article › peer-review

18 Citations (Scopus)

Abstract

Lysophosphatidic acid (LPA) stimulates Na⁺/H⁺ exchanger 3 (NHE3) activity in opossum kidney proximal tubule (OK) cells by increasing the apical membrane amount of NHE3. This occurs by stimulation of exocytic trafficking of NHE3 to the apical plasma membrane by an E3KARP-dependent mechanism. However, it is still unclear how E3KARP leads to the LPA-induced exocytosis of NHE3. In the current study, we demonstrate that stable expression of exogenous E3KARP increases LPA-induced phospholipase C (PLC) activation and subsequent elevation of intracellular Ca²⁺ in opossum kidney proximal tubule (OK) cells. Pretreatment with U73122, a PLC inhibitor, prevented the LPA-induced NHE3 activation and the exocytic trafficking of NHE3. To understand how the elevation of intracellular Ca ²⁺ leads to the stimulation of NHE3, we pretreated OK cells with BAPTA-AM, an intracellular Ca²⁺ chelator. BAPTA-AM completely blocked the LPA-induced increase of NHE3 activity and surface NHE3 amount by decreasing the LPA-induced exocytic trafficking of NHE3. Pretreatment with GF109203X, a PKC inhibitor, did not affect the percent of LPA-induced NHE3 activation and increase of surface NHE3 amount. From these results, we suggest that E3KARP plays a necessary role in LPA-induced PLC activation, and that PLC-dependent elevation of intracellular Ca²⁺ but not PKC activation is necessary for the LPA-induced increase of NHE3 exocytosis.

Original language	English
Pages (from-to)	59-68
Number of pages	10
Journal	Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids
Volume	1683
Issue number	1-3
DOIs	https://doi.org/10.1016/j.bbalip.2004.04.005
Publication status	Published - 2004 Jul 5
Externally published	Yes

Bibliographical note

Funding Information:
This work was supported in part by the Korea Research Foundation Grants (KRF-2002-070-C00074, KRF-2002-003-C00108), Pusan National University, NIH NIDDK Grants (RO1-DK26523, RO1-DK5581, RO1-DK32839, PO1-DK44484, T32 DK07632), and the Hopkins Center for Epithelial Disorders. We thank Dr. S. H. Snyder, Randon Patterson, and Byung-il Bae for their assistance with the Ca 2+ measurements.

Keywords

Calcium
E3KARP
Exocytosis
G protein-coupled receptor
GPCR
LPA
NHE3
Na/H exchanger 3
OK cells
PDZ
PLC
PSD-95/Dlg-1/ZO-1
Phospholipase C
lysophosphatidic acid
opossum kidney proximal tubule cells
phospholipase C

ASJC Scopus subject areas

Molecular Biology
Cell Biology

Access to Document

10.1016/j.bbalip.2004.04.005

Cite this

Choi, J. W., Lee-Kwon, W., Jeon, E. S., Kang, Y. J., Kawano, K., Kim, H. S., Suh, P. G., Donowitz, M., & Kim, J. H. (2004). Lysophosphatidic acid induces exocytic trafficking of Na⁺/H ⁺ exchanger 3 by E3KARP-dependent activation of phospholipase C. Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids, 1683(1-3), 59-68. https://doi.org/10.1016/j.bbalip.2004.04.005

Choi, JW, Lee-Kwon, W, Jeon, ES, Kang, YJ, Kawano, K, Kim, HS, Suh, PG, Donowitz, M & Kim, JH 2004, 'Lysophosphatidic acid induces exocytic trafficking of Na⁺/H ⁺ exchanger 3 by E3KARP-dependent activation of phospholipase C', Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids, vol. 1683, no. 1-3, pp. 59-68. https://doi.org/10.1016/j.bbalip.2004.04.005

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title = "Lysophosphatidic acid induces exocytic trafficking of Na+/H + exchanger 3 by E3KARP-dependent activation of phospholipase C",

abstract = "Lysophosphatidic acid (LPA) stimulates Na+/H+ exchanger 3 (NHE3) activity in opossum kidney proximal tubule (OK) cells by increasing the apical membrane amount of NHE3. This occurs by stimulation of exocytic trafficking of NHE3 to the apical plasma membrane by an E3KARP-dependent mechanism. However, it is still unclear how E3KARP leads to the LPA-induced exocytosis of NHE3. In the current study, we demonstrate that stable expression of exogenous E3KARP increases LPA-induced phospholipase C (PLC) activation and subsequent elevation of intracellular Ca2+ in opossum kidney proximal tubule (OK) cells. Pretreatment with U73122, a PLC inhibitor, prevented the LPA-induced NHE3 activation and the exocytic trafficking of NHE3. To understand how the elevation of intracellular Ca 2+ leads to the stimulation of NHE3, we pretreated OK cells with BAPTA-AM, an intracellular Ca2+ chelator. BAPTA-AM completely blocked the LPA-induced increase of NHE3 activity and surface NHE3 amount by decreasing the LPA-induced exocytic trafficking of NHE3. Pretreatment with GF109203X, a PKC inhibitor, did not affect the percent of LPA-induced NHE3 activation and increase of surface NHE3 amount. From these results, we suggest that E3KARP plays a necessary role in LPA-induced PLC activation, and that PLC-dependent elevation of intracellular Ca2+ but not PKC activation is necessary for the LPA-induced increase of NHE3 exocytosis.",

keywords = "Calcium, E3KARP, Exocytosis, G protein-coupled receptor, GPCR, LPA, NHE3, Na/H exchanger 3, OK cells, PDZ, PLC, PSD-95/Dlg-1/ZO-1, Phospholipase C, lysophosphatidic acid, opossum kidney proximal tubule cells, phospholipase C",

author = "Choi, {Jung Woong} and Whaseon Lee-Kwon and Jeon, {Eun Su} and Kang, {Yong Jung} and Kazuya Kawano and Kim, {Hyeon Soo} and Suh, {Pann Ghill} and Mark Donowitz and Kim, {Jae Ho}",

note = "Funding Information: This work was supported in part by the Korea Research Foundation Grants (KRF-2002-070-C00074, KRF-2002-003-C00108), Pusan National University, NIH NIDDK Grants (RO1-DK26523, RO1-DK5581, RO1-DK32839, PO1-DK44484, T32 DK07632), and the Hopkins Center for Epithelial Disorders. We thank Dr. S. H. Snyder, Randon Patterson, and Byung-il Bae for their assistance with the Ca 2+ measurements.",

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doi = "10.1016/j.bbalip.2004.04.005",

language = "English",

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pages = "59--68",

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TY - JOUR

T1 - Lysophosphatidic acid induces exocytic trafficking of Na+/H + exchanger 3 by E3KARP-dependent activation of phospholipase C

AU - Choi, Jung Woong

AU - Lee-Kwon, Whaseon

AU - Jeon, Eun Su

AU - Kang, Yong Jung

AU - Kawano, Kazuya

AU - Kim, Hyeon Soo

AU - Suh, Pann Ghill

AU - Donowitz, Mark

AU - Kim, Jae Ho

N1 - Funding Information: This work was supported in part by the Korea Research Foundation Grants (KRF-2002-070-C00074, KRF-2002-003-C00108), Pusan National University, NIH NIDDK Grants (RO1-DK26523, RO1-DK5581, RO1-DK32839, PO1-DK44484, T32 DK07632), and the Hopkins Center for Epithelial Disorders. We thank Dr. S. H. Snyder, Randon Patterson, and Byung-il Bae for their assistance with the Ca 2+ measurements.

PY - 2004/7/5

Y1 - 2004/7/5

N2 - Lysophosphatidic acid (LPA) stimulates Na+/H+ exchanger 3 (NHE3) activity in opossum kidney proximal tubule (OK) cells by increasing the apical membrane amount of NHE3. This occurs by stimulation of exocytic trafficking of NHE3 to the apical plasma membrane by an E3KARP-dependent mechanism. However, it is still unclear how E3KARP leads to the LPA-induced exocytosis of NHE3. In the current study, we demonstrate that stable expression of exogenous E3KARP increases LPA-induced phospholipase C (PLC) activation and subsequent elevation of intracellular Ca2+ in opossum kidney proximal tubule (OK) cells. Pretreatment with U73122, a PLC inhibitor, prevented the LPA-induced NHE3 activation and the exocytic trafficking of NHE3. To understand how the elevation of intracellular Ca 2+ leads to the stimulation of NHE3, we pretreated OK cells with BAPTA-AM, an intracellular Ca2+ chelator. BAPTA-AM completely blocked the LPA-induced increase of NHE3 activity and surface NHE3 amount by decreasing the LPA-induced exocytic trafficking of NHE3. Pretreatment with GF109203X, a PKC inhibitor, did not affect the percent of LPA-induced NHE3 activation and increase of surface NHE3 amount. From these results, we suggest that E3KARP plays a necessary role in LPA-induced PLC activation, and that PLC-dependent elevation of intracellular Ca2+ but not PKC activation is necessary for the LPA-induced increase of NHE3 exocytosis.

AB - Lysophosphatidic acid (LPA) stimulates Na+/H+ exchanger 3 (NHE3) activity in opossum kidney proximal tubule (OK) cells by increasing the apical membrane amount of NHE3. This occurs by stimulation of exocytic trafficking of NHE3 to the apical plasma membrane by an E3KARP-dependent mechanism. However, it is still unclear how E3KARP leads to the LPA-induced exocytosis of NHE3. In the current study, we demonstrate that stable expression of exogenous E3KARP increases LPA-induced phospholipase C (PLC) activation and subsequent elevation of intracellular Ca2+ in opossum kidney proximal tubule (OK) cells. Pretreatment with U73122, a PLC inhibitor, prevented the LPA-induced NHE3 activation and the exocytic trafficking of NHE3. To understand how the elevation of intracellular Ca 2+ leads to the stimulation of NHE3, we pretreated OK cells with BAPTA-AM, an intracellular Ca2+ chelator. BAPTA-AM completely blocked the LPA-induced increase of NHE3 activity and surface NHE3 amount by decreasing the LPA-induced exocytic trafficking of NHE3. Pretreatment with GF109203X, a PKC inhibitor, did not affect the percent of LPA-induced NHE3 activation and increase of surface NHE3 amount. From these results, we suggest that E3KARP plays a necessary role in LPA-induced PLC activation, and that PLC-dependent elevation of intracellular Ca2+ but not PKC activation is necessary for the LPA-induced increase of NHE3 exocytosis.

KW - Calcium

KW - E3KARP

KW - Exocytosis

KW - G protein-coupled receptor

KW - GPCR

KW - LPA

KW - NHE3

KW - Na/H exchanger 3

KW - OK cells

KW - PDZ

KW - PLC

KW - PSD-95/Dlg-1/ZO-1

KW - Phospholipase C

KW - lysophosphatidic acid

KW - opossum kidney proximal tubule cells

KW - phospholipase C

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DO - 10.1016/j.bbalip.2004.04.005

M3 - Article

C2 - 15238220

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SN - 1388-1981

VL - 1683

SP - 59

EP - 68

JO - Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids

JF - Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids

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