Palmitoylation of the Na/Ca exchanger cytoplasmic loop controls its inactivation and internalization during stress signaling

Louise Reilly, Jacqueline Howie, Krzysztof Wypijewski, Michael L J Ashford, Donald W. Hilgemann, William Fuller

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The electrogenicNa/Ca exchanger (NCX) mediates bidirectional Ca movements that are highly sensitive to changes of Na gradients in many cells. NCX1 is implicated in the pathogenesis of heart failure and a number of cardiac arrhythmias. We measured NCX1 palmitoylation using resin-assisted capture, the subcellular location of yellow fluorescent protein-NCX1 fusion proteins, and NCX1 currents using whole-cell voltage clamping. Rat NCX1 is substantially palmitoylated in all tissues examined. Cysteine 739 in the NCX1 large intracellular loop is necessary and sufficient for NCX1 palmitoylation. Palmitoylation ofNCX1occurs in the Golgi and anchors theNCX1 large regulatory intracellular loop to membranes. Surprisingly, palmitoylation does not influence trafficking or localization of NCX1 to surface membranes, nor does it strongly affect the normal forward or reverse transport modes of NCX1. However, exchangers that cannot be palmitoylated do not inactivate normally (leading to substantial activity in conditions when wild-type exchangers are inactive) and do not promote cargo-dependent endocytosis that internalizes 50% of the cell surface following strong G-protein activation or largeCa transients. The palmitoylated cysteine inNCX1 is found in all vertebrate and some invertebrate NCX homologs. Thus, NCX palmitoylation ubiquitously modulates Ca homeostasis and membrane domain function in cells that expressNCXproteins. - Reilly,L.,Howie, J.,Wypijewski, K., Ashford,M.L. J.,Hilgemann, D. W., Fuller,W.Palmitoylation of the Na/Ca exchanger cytoplasmic loop controls its inactivation and internalization during stress signaling.

Original languageEnglish (US)
Pages (from-to)4532-4543
Number of pages12
JournalFASEB Journal
Volume29
Issue number11
DOIs
StatePublished - Nov 1 2015

Fingerprint

Lipoylation
Membranes
Cysteine
Anchors
GTP-Binding Proteins
Rats
Fusion reactions
Resins
Chemical activation
Tissue
Electric potential
Invertebrates
Endocytosis
Constriction
Vertebrates
Cardiac Arrhythmias
Homeostasis
Heart Failure
sodium-calcium exchanger 1

Keywords

  • Acylation
  • Heart
  • Ion transport
  • Patch clamp
  • Resin-assisted capture

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Genetics
  • Molecular Biology

Cite this

Palmitoylation of the Na/Ca exchanger cytoplasmic loop controls its inactivation and internalization during stress signaling. / Reilly, Louise; Howie, Jacqueline; Wypijewski, Krzysztof; Ashford, Michael L J; Hilgemann, Donald W.; Fuller, William.

In: FASEB Journal, Vol. 29, No. 11, 01.11.2015, p. 4532-4543.

Research output: Contribution to journalArticle

Reilly, Louise ; Howie, Jacqueline ; Wypijewski, Krzysztof ; Ashford, Michael L J ; Hilgemann, Donald W. ; Fuller, William. / Palmitoylation of the Na/Ca exchanger cytoplasmic loop controls its inactivation and internalization during stress signaling. In: FASEB Journal. 2015 ; Vol. 29, No. 11. pp. 4532-4543.
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N2 - The electrogenicNa/Ca exchanger (NCX) mediates bidirectional Ca movements that are highly sensitive to changes of Na gradients in many cells. NCX1 is implicated in the pathogenesis of heart failure and a number of cardiac arrhythmias. We measured NCX1 palmitoylation using resin-assisted capture, the subcellular location of yellow fluorescent protein-NCX1 fusion proteins, and NCX1 currents using whole-cell voltage clamping. Rat NCX1 is substantially palmitoylated in all tissues examined. Cysteine 739 in the NCX1 large intracellular loop is necessary and sufficient for NCX1 palmitoylation. Palmitoylation ofNCX1occurs in the Golgi and anchors theNCX1 large regulatory intracellular loop to membranes. Surprisingly, palmitoylation does not influence trafficking or localization of NCX1 to surface membranes, nor does it strongly affect the normal forward or reverse transport modes of NCX1. However, exchangers that cannot be palmitoylated do not inactivate normally (leading to substantial activity in conditions when wild-type exchangers are inactive) and do not promote cargo-dependent endocytosis that internalizes 50% of the cell surface following strong G-protein activation or largeCa transients. The palmitoylated cysteine inNCX1 is found in all vertebrate and some invertebrate NCX homologs. Thus, NCX palmitoylation ubiquitously modulates Ca homeostasis and membrane domain function in cells that expressNCXproteins. - Reilly,L.,Howie, J.,Wypijewski, K., Ashford,M.L. J.,Hilgemann, D. W., Fuller,W.Palmitoylation of the Na/Ca exchanger cytoplasmic loop controls its inactivation and internalization during stress signaling.

AB - The electrogenicNa/Ca exchanger (NCX) mediates bidirectional Ca movements that are highly sensitive to changes of Na gradients in many cells. NCX1 is implicated in the pathogenesis of heart failure and a number of cardiac arrhythmias. We measured NCX1 palmitoylation using resin-assisted capture, the subcellular location of yellow fluorescent protein-NCX1 fusion proteins, and NCX1 currents using whole-cell voltage clamping. Rat NCX1 is substantially palmitoylated in all tissues examined. Cysteine 739 in the NCX1 large intracellular loop is necessary and sufficient for NCX1 palmitoylation. Palmitoylation ofNCX1occurs in the Golgi and anchors theNCX1 large regulatory intracellular loop to membranes. Surprisingly, palmitoylation does not influence trafficking or localization of NCX1 to surface membranes, nor does it strongly affect the normal forward or reverse transport modes of NCX1. However, exchangers that cannot be palmitoylated do not inactivate normally (leading to substantial activity in conditions when wild-type exchangers are inactive) and do not promote cargo-dependent endocytosis that internalizes 50% of the cell surface following strong G-protein activation or largeCa transients. The palmitoylated cysteine inNCX1 is found in all vertebrate and some invertebrate NCX homologs. Thus, NCX palmitoylation ubiquitously modulates Ca homeostasis and membrane domain function in cells that expressNCXproteins. - Reilly,L.,Howie, J.,Wypijewski, K., Ashford,M.L. J.,Hilgemann, D. W., Fuller,W.Palmitoylation of the Na/Ca exchanger cytoplasmic loop controls its inactivation and internalization during stress signaling.

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