Phospholipase C-γ binds directly to the Na+/H+ exchanger 3 and is required for calcium regulation of exchange activity

Nicholas C. Zachos, Damian B. van Rossum, Xuhang Li, Gabriela Caraveo, Rafiquel Sarker, Boyoung Cha, Sachin Mohan, Stephen Desiderio, Randen L. Patterson, Mark Donowitz

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Multiple studies suggest that phospholipase C-γ(PLC-γ) contributes to regulation of sodium/hydrogen exchanger 3 (NHE3) in the small intestine, although the mechanism(s) for this regulation remain unknown. We demonstrate here that PLC-γ binds directly to the C terminus of NHE3 and exists in similar sized multiprotein complexes as NHE3. This binding is dynamic and decreases with elevated [Ca2+]i. The PLC-γ-binding site in NHE3 was identified (amino acids 586-605) and shown to be a critical regulatory domain for protein complex formation, because when it is mutated, NHE3 binding to PLC-γ as well as NHERF2 is lost. An inhibitory peptide, which binds to the Src homology 2 domains contained in PLC-γ without interrupting binding of PLC-γ to NHE3, was used to probe a non-lipase-dependent role of PLC-γ. In the presence of this peptide, carbachol- stimulated calcium inhibition of NHE3 was lost. These results mirror previous studies with the transient receptor potential channel and suggest that PLC-γ may play a common role in regulating the cell-surface expression of ion transporters.

Original languageEnglish (US)
Pages (from-to)19437-19444
Number of pages8
JournalJournal of Biological Chemistry
Volume284
Issue number29
DOIs
StatePublished - Jul 17 2009
Externally publishedYes

Fingerprint

Sodium-Hydrogen Antiporter
Type C Phospholipases
Programmable logic controllers
Calcium
Transient Receptor Potential Channels
Multiprotein Complexes
Peptides
src Homology Domains
Carbachol
Small Intestine
Binding Sites
Ions
Amino Acids
Mirrors
sodium-hydrogen exchanger 3
Protein Domains

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Zachos, N. C., van Rossum, D. B., Li, X., Caraveo, G., Sarker, R., Cha, B., ... Donowitz, M. (2009). Phospholipase C-γ binds directly to the Na+/H+ exchanger 3 and is required for calcium regulation of exchange activity. Journal of Biological Chemistry, 284(29), 19437-19444. https://doi.org/10.1074/jbc.M109.006098

Phospholipase C-γ binds directly to the Na+/H+ exchanger 3 and is required for calcium regulation of exchange activity. / Zachos, Nicholas C.; van Rossum, Damian B.; Li, Xuhang; Caraveo, Gabriela; Sarker, Rafiquel; Cha, Boyoung; Mohan, Sachin; Desiderio, Stephen; Patterson, Randen L.; Donowitz, Mark.

In: Journal of Biological Chemistry, Vol. 284, No. 29, 17.07.2009, p. 19437-19444.

Research output: Contribution to journalArticle

Zachos, NC, van Rossum, DB, Li, X, Caraveo, G, Sarker, R, Cha, B, Mohan, S, Desiderio, S, Patterson, RL & Donowitz, M 2009, 'Phospholipase C-γ binds directly to the Na+/H+ exchanger 3 and is required for calcium regulation of exchange activity', Journal of Biological Chemistry, vol. 284, no. 29, pp. 19437-19444. https://doi.org/10.1074/jbc.M109.006098
Zachos, Nicholas C. ; van Rossum, Damian B. ; Li, Xuhang ; Caraveo, Gabriela ; Sarker, Rafiquel ; Cha, Boyoung ; Mohan, Sachin ; Desiderio, Stephen ; Patterson, Randen L. ; Donowitz, Mark. / Phospholipase C-γ binds directly to the Na+/H+ exchanger 3 and is required for calcium regulation of exchange activity. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 29. pp. 19437-19444.
@article{1c009d82ea4747f7a0ce8496b4756f9b,
title = "Phospholipase C-γ binds directly to the Na+/H+ exchanger 3 and is required for calcium regulation of exchange activity",
abstract = "Multiple studies suggest that phospholipase C-γ(PLC-γ) contributes to regulation of sodium/hydrogen exchanger 3 (NHE3) in the small intestine, although the mechanism(s) for this regulation remain unknown. We demonstrate here that PLC-γ binds directly to the C terminus of NHE3 and exists in similar sized multiprotein complexes as NHE3. This binding is dynamic and decreases with elevated [Ca2+]i. The PLC-γ-binding site in NHE3 was identified (amino acids 586-605) and shown to be a critical regulatory domain for protein complex formation, because when it is mutated, NHE3 binding to PLC-γ as well as NHERF2 is lost. An inhibitory peptide, which binds to the Src homology 2 domains contained in PLC-γ without interrupting binding of PLC-γ to NHE3, was used to probe a non-lipase-dependent role of PLC-γ. In the presence of this peptide, carbachol- stimulated calcium inhibition of NHE3 was lost. These results mirror previous studies with the transient receptor potential channel and suggest that PLC-γ may play a common role in regulating the cell-surface expression of ion transporters.",
author = "Zachos, {Nicholas C.} and {van Rossum}, {Damian B.} and Xuhang Li and Gabriela Caraveo and Rafiquel Sarker and Boyoung Cha and Sachin Mohan and Stephen Desiderio and Patterson, {Randen L.} and Mark Donowitz",
year = "2009",
month = "7",
day = "17",
doi = "10.1074/jbc.M109.006098",
language = "English (US)",
volume = "284",
pages = "19437--19444",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "29",

}

TY - JOUR

T1 - Phospholipase C-γ binds directly to the Na+/H+ exchanger 3 and is required for calcium regulation of exchange activity

AU - Zachos, Nicholas C.

AU - van Rossum, Damian B.

AU - Li, Xuhang

AU - Caraveo, Gabriela

AU - Sarker, Rafiquel

AU - Cha, Boyoung

AU - Mohan, Sachin

AU - Desiderio, Stephen

AU - Patterson, Randen L.

AU - Donowitz, Mark

PY - 2009/7/17

Y1 - 2009/7/17

N2 - Multiple studies suggest that phospholipase C-γ(PLC-γ) contributes to regulation of sodium/hydrogen exchanger 3 (NHE3) in the small intestine, although the mechanism(s) for this regulation remain unknown. We demonstrate here that PLC-γ binds directly to the C terminus of NHE3 and exists in similar sized multiprotein complexes as NHE3. This binding is dynamic and decreases with elevated [Ca2+]i. The PLC-γ-binding site in NHE3 was identified (amino acids 586-605) and shown to be a critical regulatory domain for protein complex formation, because when it is mutated, NHE3 binding to PLC-γ as well as NHERF2 is lost. An inhibitory peptide, which binds to the Src homology 2 domains contained in PLC-γ without interrupting binding of PLC-γ to NHE3, was used to probe a non-lipase-dependent role of PLC-γ. In the presence of this peptide, carbachol- stimulated calcium inhibition of NHE3 was lost. These results mirror previous studies with the transient receptor potential channel and suggest that PLC-γ may play a common role in regulating the cell-surface expression of ion transporters.

AB - Multiple studies suggest that phospholipase C-γ(PLC-γ) contributes to regulation of sodium/hydrogen exchanger 3 (NHE3) in the small intestine, although the mechanism(s) for this regulation remain unknown. We demonstrate here that PLC-γ binds directly to the C terminus of NHE3 and exists in similar sized multiprotein complexes as NHE3. This binding is dynamic and decreases with elevated [Ca2+]i. The PLC-γ-binding site in NHE3 was identified (amino acids 586-605) and shown to be a critical regulatory domain for protein complex formation, because when it is mutated, NHE3 binding to PLC-γ as well as NHERF2 is lost. An inhibitory peptide, which binds to the Src homology 2 domains contained in PLC-γ without interrupting binding of PLC-γ to NHE3, was used to probe a non-lipase-dependent role of PLC-γ. In the presence of this peptide, carbachol- stimulated calcium inhibition of NHE3 was lost. These results mirror previous studies with the transient receptor potential channel and suggest that PLC-γ may play a common role in regulating the cell-surface expression of ion transporters.

UR - http://www.scopus.com/inward/record.url?scp=67749096203&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67749096203&partnerID=8YFLogxK

U2 - 10.1074/jbc.M109.006098

DO - 10.1074/jbc.M109.006098

M3 - Article

C2 - 19473983

AN - SCOPUS:67749096203

VL - 284

SP - 19437

EP - 19444

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 29

ER -