Caveolin-1 assembles type 1 inositol 1,4,5-trisphosphate receptors and canonical transient receptor potential 3 channels into a functional signaling complex in arterial smooth muscle cells

Adebowale Adebiyi, Damodaran Narayanan, Jonathan Jaggar

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Abstract

Physical coupling of sarcoplasmic reticulum (SR) type 1 inositol 1,4,5-trisphosphate receptors (IP3R1) to plasma membrane canonical transient receptor potential 3 (TRPC3) channels activates a cation current (ICat) in arterial smooth muscle cells that induces vasoconstriction. However, structural components that enable IP3R1 and TRPC3 channels to communicate locally are unclear. Caveolae are plasma membrane microdomains that can compartmentalize proteins. Here, we tested the hypothesis that caveolae and specifically caveolin-1 (cav-1), a caveolae scaffolding protein, facilitate functional IP3R1 to TRPC3 coupling in smooth muscle cells of resistance-size cerebral arteries. Methyl-β-cyclodextrin (MβCD), which disassembles caveolae, reduced IP3-induced ICat activation in smooth muscle cells and vasoconstriction in pressurized arteries. Cholesterol replenishment reversed these effects. Cav-1 knockdown using shRNA attenuated IP3-induced vasoconstriction, but did not alter TRPC3 and IP3R1 expression. A synthetic peptide corresponding to the cav-1 scaffolding domain (CSD) sequence (amino acids 82-101) also attenuated IP 3-induced ICat activation and vasoconstriction. A cav-1 antibody co-immunoprecipitated cav-1, TRPC3, and IP3R1 from cerebral artery lysate. ImmunoFRET indicated that cav-1, TRPC3 channels and IP 3R1 are spatially co-localized in arterial smooth muscle cells. IP3R1 and TRPC3 channel spatial localization was disrupted by MβCD and a CSD peptide. Cholesterol replenishment re-established IP3R1 and TRPC3 channel close spatial proximity. Taken together, these data indicate that in arterial smooth muscle cells, cav-1 co-localizes SR IP3R1 and plasma membrane TRPC3 channels in close spatial proximity thereby enabling IP 3-induced physical coupling of these proteins, leading to I Cat generation and vasoconstriction.

Original languageEnglish (US)
Pages (from-to)4341-4348
Number of pages8
JournalJournal of Biological Chemistry
Volume286
Issue number6
DOIs
StatePublished - Feb 11 2011

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Transient Receptor Potential Channels
Inositol 1,4,5-Trisphosphate Receptors
Caveolin 1
Smooth Muscle Myocytes
Muscle
Caveolae
Vasoconstriction
Cells
Cell membranes
Cerebral Arteries
Cell Membrane
Cyclodextrins
Sarcoplasmic Reticulum
Caveolin 3
Chemical activation
Cholesterol
Membrane Microdomains
Peptides
Proteins
Ion Channels

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

@article{3d1408ef58db4ff193bdfc9ef9404fbd,
title = "Caveolin-1 assembles type 1 inositol 1,4,5-trisphosphate receptors and canonical transient receptor potential 3 channels into a functional signaling complex in arterial smooth muscle cells",
abstract = "Physical coupling of sarcoplasmic reticulum (SR) type 1 inositol 1,4,5-trisphosphate receptors (IP3R1) to plasma membrane canonical transient receptor potential 3 (TRPC3) channels activates a cation current (ICat) in arterial smooth muscle cells that induces vasoconstriction. However, structural components that enable IP3R1 and TRPC3 channels to communicate locally are unclear. Caveolae are plasma membrane microdomains that can compartmentalize proteins. Here, we tested the hypothesis that caveolae and specifically caveolin-1 (cav-1), a caveolae scaffolding protein, facilitate functional IP3R1 to TRPC3 coupling in smooth muscle cells of resistance-size cerebral arteries. Methyl-β-cyclodextrin (MβCD), which disassembles caveolae, reduced IP3-induced ICat activation in smooth muscle cells and vasoconstriction in pressurized arteries. Cholesterol replenishment reversed these effects. Cav-1 knockdown using shRNA attenuated IP3-induced vasoconstriction, but did not alter TRPC3 and IP3R1 expression. A synthetic peptide corresponding to the cav-1 scaffolding domain (CSD) sequence (amino acids 82-101) also attenuated IP 3-induced ICat activation and vasoconstriction. A cav-1 antibody co-immunoprecipitated cav-1, TRPC3, and IP3R1 from cerebral artery lysate. ImmunoFRET indicated that cav-1, TRPC3 channels and IP 3R1 are spatially co-localized in arterial smooth muscle cells. IP3R1 and TRPC3 channel spatial localization was disrupted by MβCD and a CSD peptide. Cholesterol replenishment re-established IP3R1 and TRPC3 channel close spatial proximity. Taken together, these data indicate that in arterial smooth muscle cells, cav-1 co-localizes SR IP3R1 and plasma membrane TRPC3 channels in close spatial proximity thereby enabling IP 3-induced physical coupling of these proteins, leading to I Cat generation and vasoconstriction.",
author = "Adebowale Adebiyi and Damodaran Narayanan and Jonathan Jaggar",
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T1 - Caveolin-1 assembles type 1 inositol 1,4,5-trisphosphate receptors and canonical transient receptor potential 3 channels into a functional signaling complex in arterial smooth muscle cells

AU - Adebiyi, Adebowale

AU - Narayanan, Damodaran

AU - Jaggar, Jonathan

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N2 - Physical coupling of sarcoplasmic reticulum (SR) type 1 inositol 1,4,5-trisphosphate receptors (IP3R1) to plasma membrane canonical transient receptor potential 3 (TRPC3) channels activates a cation current (ICat) in arterial smooth muscle cells that induces vasoconstriction. However, structural components that enable IP3R1 and TRPC3 channels to communicate locally are unclear. Caveolae are plasma membrane microdomains that can compartmentalize proteins. Here, we tested the hypothesis that caveolae and specifically caveolin-1 (cav-1), a caveolae scaffolding protein, facilitate functional IP3R1 to TRPC3 coupling in smooth muscle cells of resistance-size cerebral arteries. Methyl-β-cyclodextrin (MβCD), which disassembles caveolae, reduced IP3-induced ICat activation in smooth muscle cells and vasoconstriction in pressurized arteries. Cholesterol replenishment reversed these effects. Cav-1 knockdown using shRNA attenuated IP3-induced vasoconstriction, but did not alter TRPC3 and IP3R1 expression. A synthetic peptide corresponding to the cav-1 scaffolding domain (CSD) sequence (amino acids 82-101) also attenuated IP 3-induced ICat activation and vasoconstriction. A cav-1 antibody co-immunoprecipitated cav-1, TRPC3, and IP3R1 from cerebral artery lysate. ImmunoFRET indicated that cav-1, TRPC3 channels and IP 3R1 are spatially co-localized in arterial smooth muscle cells. IP3R1 and TRPC3 channel spatial localization was disrupted by MβCD and a CSD peptide. Cholesterol replenishment re-established IP3R1 and TRPC3 channel close spatial proximity. Taken together, these data indicate that in arterial smooth muscle cells, cav-1 co-localizes SR IP3R1 and plasma membrane TRPC3 channels in close spatial proximity thereby enabling IP 3-induced physical coupling of these proteins, leading to I Cat generation and vasoconstriction.

AB - Physical coupling of sarcoplasmic reticulum (SR) type 1 inositol 1,4,5-trisphosphate receptors (IP3R1) to plasma membrane canonical transient receptor potential 3 (TRPC3) channels activates a cation current (ICat) in arterial smooth muscle cells that induces vasoconstriction. However, structural components that enable IP3R1 and TRPC3 channels to communicate locally are unclear. Caveolae are plasma membrane microdomains that can compartmentalize proteins. Here, we tested the hypothesis that caveolae and specifically caveolin-1 (cav-1), a caveolae scaffolding protein, facilitate functional IP3R1 to TRPC3 coupling in smooth muscle cells of resistance-size cerebral arteries. Methyl-β-cyclodextrin (MβCD), which disassembles caveolae, reduced IP3-induced ICat activation in smooth muscle cells and vasoconstriction in pressurized arteries. Cholesterol replenishment reversed these effects. Cav-1 knockdown using shRNA attenuated IP3-induced vasoconstriction, but did not alter TRPC3 and IP3R1 expression. A synthetic peptide corresponding to the cav-1 scaffolding domain (CSD) sequence (amino acids 82-101) also attenuated IP 3-induced ICat activation and vasoconstriction. A cav-1 antibody co-immunoprecipitated cav-1, TRPC3, and IP3R1 from cerebral artery lysate. ImmunoFRET indicated that cav-1, TRPC3 channels and IP 3R1 are spatially co-localized in arterial smooth muscle cells. IP3R1 and TRPC3 channel spatial localization was disrupted by MβCD and a CSD peptide. Cholesterol replenishment re-established IP3R1 and TRPC3 channel close spatial proximity. Taken together, these data indicate that in arterial smooth muscle cells, cav-1 co-localizes SR IP3R1 and plasma membrane TRPC3 channels in close spatial proximity thereby enabling IP 3-induced physical coupling of these proteins, leading to I Cat generation and vasoconstriction.

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