LRRC26 is a functional BK channel auxiliary γ subunit in arterial smooth muscle cells

Kirk W. Evanson, John P. Bannister, Marie Dennis Leo, Jonathan Jaggar

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

RATIONALE: Smooth muscle cell (myocyte) large-conductance calcium (Ca)-activated potassium (BK) channels are functionally significant modulators of arterial contractility. Arterial myocytes express both pore-forming BKα and auxiliary β1 subunits, which increase channel Ca sensitivity. Recently, several leucine-rich repeat containing (LRRC) proteins have been identified as auxiliary γ subunits that elevate the voltage sensitivity of recombinant and prostate adenocarcinoma BK channels. LRRC expression and physiological functions in native cell types are unclear. OBJECTIVE:: Investigate the expression and physiological functions of leucine-rich repeat containing protein 26 (LRRC26) in arterial myocytes. METHODS AND RESULTS:: Reverse transcription polymerase chain reaction and Western blotting detected LRRC26 mRNA and protein in cerebral artery myocytes. Biotinylation, immunofluorescence resonance energy transfer microscopy, and coimmunoprecipitation indicated that LRRC26 was located in close spatial proximity to, and associated with, plasma membrane BKα subunits. LRRC26 knockdown (RNAi) reduced total and surface LRRC26, but did not alter BKα or β1, proteins in arteries. LRRC26 knockdown did not alter Ca sparks but reduced BK channel voltage sensitivity, which decreased channel apparent Ca sensitivity and transient BK current frequency and amplitude in myocytes. LRRC26 knockdown also increased myogenic tone over a range (40-100 mm Hg) of intravascular pressures, and reduced vasoconstriction to iberiotoxin and vasodilation to NS1619, BK channel inhibitors and activators, respectively. In contrast, LRRC26 knockdown did not alter depolarization (60 mmol/L K)-induced vasoconstriction. CONCLUSIONS:: LRRC26 is expressed, associates with BKα subunits, and elevates channel voltage- and apparent Ca sensitivity in arterial myocytes to induce vasodilation. This study indicates that arterial myocytes express a functional BK channel γ subunit.

Original languageEnglish (US)
Pages (from-to)423-431
Number of pages9
JournalCirculation research
Volume115
Issue number4
DOIs
StatePublished - Aug 1 2014

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Large-Conductance Calcium-Activated Potassium Channels
Smooth Muscle Myocytes
Muscle Cells
Calcium Channels
Vasoconstriction
Vasodilation
leucine-rich repeat proteins
Biotinylation
Cerebral Arteries
Calcium Signaling
Energy Transfer
RNA Interference
Leucine
Reverse Transcription
Fluorescent Antibody Technique
Prostate
Microscopy
Proteins
Adenocarcinoma
Arteries

All Science Journal Classification (ASJC) codes

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

LRRC26 is a functional BK channel auxiliary γ subunit in arterial smooth muscle cells. / Evanson, Kirk W.; Bannister, John P.; Leo, Marie Dennis; Jaggar, Jonathan.

In: Circulation research, Vol. 115, No. 4, 01.08.2014, p. 423-431.

Research output: Contribution to journalArticle

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abstract = "RATIONALE: Smooth muscle cell (myocyte) large-conductance calcium (Ca)-activated potassium (BK) channels are functionally significant modulators of arterial contractility. Arterial myocytes express both pore-forming BKα and auxiliary β1 subunits, which increase channel Ca sensitivity. Recently, several leucine-rich repeat containing (LRRC) proteins have been identified as auxiliary γ subunits that elevate the voltage sensitivity of recombinant and prostate adenocarcinoma BK channels. LRRC expression and physiological functions in native cell types are unclear. OBJECTIVE:: Investigate the expression and physiological functions of leucine-rich repeat containing protein 26 (LRRC26) in arterial myocytes. METHODS AND RESULTS:: Reverse transcription polymerase chain reaction and Western blotting detected LRRC26 mRNA and protein in cerebral artery myocytes. Biotinylation, immunofluorescence resonance energy transfer microscopy, and coimmunoprecipitation indicated that LRRC26 was located in close spatial proximity to, and associated with, plasma membrane BKα subunits. LRRC26 knockdown (RNAi) reduced total and surface LRRC26, but did not alter BKα or β1, proteins in arteries. LRRC26 knockdown did not alter Ca sparks but reduced BK channel voltage sensitivity, which decreased channel apparent Ca sensitivity and transient BK current frequency and amplitude in myocytes. LRRC26 knockdown also increased myogenic tone over a range (40-100 mm Hg) of intravascular pressures, and reduced vasoconstriction to iberiotoxin and vasodilation to NS1619, BK channel inhibitors and activators, respectively. In contrast, LRRC26 knockdown did not alter depolarization (60 mmol/L K)-induced vasoconstriction. CONCLUSIONS:: LRRC26 is expressed, associates with BKα subunits, and elevates channel voltage- and apparent Ca sensitivity in arterial myocytes to induce vasodilation. This study indicates that arterial myocytes express a functional BK channel γ subunit.",
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AU - Bannister, John P.

AU - Leo, Marie Dennis

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N2 - RATIONALE: Smooth muscle cell (myocyte) large-conductance calcium (Ca)-activated potassium (BK) channels are functionally significant modulators of arterial contractility. Arterial myocytes express both pore-forming BKα and auxiliary β1 subunits, which increase channel Ca sensitivity. Recently, several leucine-rich repeat containing (LRRC) proteins have been identified as auxiliary γ subunits that elevate the voltage sensitivity of recombinant and prostate adenocarcinoma BK channels. LRRC expression and physiological functions in native cell types are unclear. OBJECTIVE:: Investigate the expression and physiological functions of leucine-rich repeat containing protein 26 (LRRC26) in arterial myocytes. METHODS AND RESULTS:: Reverse transcription polymerase chain reaction and Western blotting detected LRRC26 mRNA and protein in cerebral artery myocytes. Biotinylation, immunofluorescence resonance energy transfer microscopy, and coimmunoprecipitation indicated that LRRC26 was located in close spatial proximity to, and associated with, plasma membrane BKα subunits. LRRC26 knockdown (RNAi) reduced total and surface LRRC26, but did not alter BKα or β1, proteins in arteries. LRRC26 knockdown did not alter Ca sparks but reduced BK channel voltage sensitivity, which decreased channel apparent Ca sensitivity and transient BK current frequency and amplitude in myocytes. LRRC26 knockdown also increased myogenic tone over a range (40-100 mm Hg) of intravascular pressures, and reduced vasoconstriction to iberiotoxin and vasodilation to NS1619, BK channel inhibitors and activators, respectively. In contrast, LRRC26 knockdown did not alter depolarization (60 mmol/L K)-induced vasoconstriction. CONCLUSIONS:: LRRC26 is expressed, associates with BKα subunits, and elevates channel voltage- and apparent Ca sensitivity in arterial myocytes to induce vasodilation. This study indicates that arterial myocytes express a functional BK channel γ subunit.

AB - RATIONALE: Smooth muscle cell (myocyte) large-conductance calcium (Ca)-activated potassium (BK) channels are functionally significant modulators of arterial contractility. Arterial myocytes express both pore-forming BKα and auxiliary β1 subunits, which increase channel Ca sensitivity. Recently, several leucine-rich repeat containing (LRRC) proteins have been identified as auxiliary γ subunits that elevate the voltage sensitivity of recombinant and prostate adenocarcinoma BK channels. LRRC expression and physiological functions in native cell types are unclear. OBJECTIVE:: Investigate the expression and physiological functions of leucine-rich repeat containing protein 26 (LRRC26) in arterial myocytes. METHODS AND RESULTS:: Reverse transcription polymerase chain reaction and Western blotting detected LRRC26 mRNA and protein in cerebral artery myocytes. Biotinylation, immunofluorescence resonance energy transfer microscopy, and coimmunoprecipitation indicated that LRRC26 was located in close spatial proximity to, and associated with, plasma membrane BKα subunits. LRRC26 knockdown (RNAi) reduced total and surface LRRC26, but did not alter BKα or β1, proteins in arteries. LRRC26 knockdown did not alter Ca sparks but reduced BK channel voltage sensitivity, which decreased channel apparent Ca sensitivity and transient BK current frequency and amplitude in myocytes. LRRC26 knockdown also increased myogenic tone over a range (40-100 mm Hg) of intravascular pressures, and reduced vasoconstriction to iberiotoxin and vasodilation to NS1619, BK channel inhibitors and activators, respectively. In contrast, LRRC26 knockdown did not alter depolarization (60 mmol/L K)-induced vasoconstriction. CONCLUSIONS:: LRRC26 is expressed, associates with BKα subunits, and elevates channel voltage- and apparent Ca sensitivity in arterial myocytes to induce vasodilation. This study indicates that arterial myocytes express a functional BK channel γ subunit.

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