Role of hβ1 in activation of human mesangial BK channels by cGMP kinase

Patrick E. Kudlacek, Jennifer L. Pluznick, Rong Ma, Babu Padanilam, Steven C. Sansom

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

In vascular smooth muscle and glomerular mesangial cells, relaxing agents such as nitric oxide and atrial natriuretic peptide activate large - conductance Ca2+-activated K+ channels (BK) via the cGMP kinase pathway. BK are composed of pore-forming α-subunits, encoded by the slopoke gene (Slo), and one of four cell-specific accessory β-subunits (hβ1-4). We used patch-clamp analysis to determine the influence of hβ1, hβ2, and hβ4 on activation of human mesangial BK by cGMP kinase. We found that HEK 293 cells, coexpressing human (h) Sloα with either hβ1 or hβ2, contained single BK currents activated by db-cGMP in cell-attached patches. However, recombinant BK were not activated by db-cGMP when hSloα was expressed alone or with hβ4. DNA-RNA hybridization revealed that mesangial cells contained mRNA for hβ1 but not hβ2 or hβ4. The BK response to db-cGMP was decreased when hβ1 antisense but not scrambled oligonucleotides were incorporated into mesangial cells. Western blot analysis showed that hβ1 antisense oligonucleotide inhibited the amount of hβ1-V5 fusion protein expressed in HEK 293 cells by ∼50%. These results show that mesangial cells contain hβ1, a BK accessory protein, which confers activation of BK by cGMP kinase.

Original languageEnglish
Pages (from-to)F289-F294
JournalAmerican Journal of Physiology - Renal Physiology
Volume285
Issue number2 54-2
StatePublished - 1 Aug 2003

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Large-Conductance Calcium-Activated Potassium Channels
Mesangial Cells
Phosphotransferases
HEK293 Cells
Calcium-Activated Potassium Channels
Antisense Oligonucleotides
Atrial Natriuretic Factor
Vascular Smooth Muscle
Oligonucleotides
Nitric Oxide
Proteins
Western Blotting
RNA
Messenger RNA
DNA
Genes

Keywords

  • Antisense
  • Guanosine 3′,5′-cyclic monophosphate
  • Human β-subunit
  • Large-conductance calcium-activated potassium channels
  • Maxi-potassium
  • Patch clamp

Cite this

Kudlacek, P. E., Pluznick, J. L., Ma, R., Padanilam, B., & Sansom, S. C. (2003). Role of hβ1 in activation of human mesangial BK channels by cGMP kinase. American Journal of Physiology - Renal Physiology, 285(2 54-2), F289-F294.
Kudlacek, Patrick E. ; Pluznick, Jennifer L. ; Ma, Rong ; Padanilam, Babu ; Sansom, Steven C. / Role of hβ1 in activation of human mesangial BK channels by cGMP kinase. In: American Journal of Physiology - Renal Physiology. 2003 ; Vol. 285, No. 2 54-2. pp. F289-F294.
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Kudlacek, PE, Pluznick, JL, Ma, R, Padanilam, B & Sansom, SC 2003, 'Role of hβ1 in activation of human mesangial BK channels by cGMP kinase', American Journal of Physiology - Renal Physiology, vol. 285, no. 2 54-2, pp. F289-F294.

Role of hβ1 in activation of human mesangial BK channels by cGMP kinase. / Kudlacek, Patrick E.; Pluznick, Jennifer L.; Ma, Rong; Padanilam, Babu; Sansom, Steven C.

In: American Journal of Physiology - Renal Physiology, Vol. 285, No. 2 54-2, 01.08.2003, p. F289-F294.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Role of hβ1 in activation of human mesangial BK channels by cGMP kinase

AU - Kudlacek, Patrick E.

AU - Pluznick, Jennifer L.

AU - Ma, Rong

AU - Padanilam, Babu

AU - Sansom, Steven C.

PY - 2003/8/1

Y1 - 2003/8/1

N2 - In vascular smooth muscle and glomerular mesangial cells, relaxing agents such as nitric oxide and atrial natriuretic peptide activate large - conductance Ca2+-activated K+ channels (BK) via the cGMP kinase pathway. BK are composed of pore-forming α-subunits, encoded by the slopoke gene (Slo), and one of four cell-specific accessory β-subunits (hβ1-4). We used patch-clamp analysis to determine the influence of hβ1, hβ2, and hβ4 on activation of human mesangial BK by cGMP kinase. We found that HEK 293 cells, coexpressing human (h) Sloα with either hβ1 or hβ2, contained single BK currents activated by db-cGMP in cell-attached patches. However, recombinant BK were not activated by db-cGMP when hSloα was expressed alone or with hβ4. DNA-RNA hybridization revealed that mesangial cells contained mRNA for hβ1 but not hβ2 or hβ4. The BK response to db-cGMP was decreased when hβ1 antisense but not scrambled oligonucleotides were incorporated into mesangial cells. Western blot analysis showed that hβ1 antisense oligonucleotide inhibited the amount of hβ1-V5 fusion protein expressed in HEK 293 cells by ∼50%. These results show that mesangial cells contain hβ1, a BK accessory protein, which confers activation of BK by cGMP kinase.

AB - In vascular smooth muscle and glomerular mesangial cells, relaxing agents such as nitric oxide and atrial natriuretic peptide activate large - conductance Ca2+-activated K+ channels (BK) via the cGMP kinase pathway. BK are composed of pore-forming α-subunits, encoded by the slopoke gene (Slo), and one of four cell-specific accessory β-subunits (hβ1-4). We used patch-clamp analysis to determine the influence of hβ1, hβ2, and hβ4 on activation of human mesangial BK by cGMP kinase. We found that HEK 293 cells, coexpressing human (h) Sloα with either hβ1 or hβ2, contained single BK currents activated by db-cGMP in cell-attached patches. However, recombinant BK were not activated by db-cGMP when hSloα was expressed alone or with hβ4. DNA-RNA hybridization revealed that mesangial cells contained mRNA for hβ1 but not hβ2 or hβ4. The BK response to db-cGMP was decreased when hβ1 antisense but not scrambled oligonucleotides were incorporated into mesangial cells. Western blot analysis showed that hβ1 antisense oligonucleotide inhibited the amount of hβ1-V5 fusion protein expressed in HEK 293 cells by ∼50%. These results show that mesangial cells contain hβ1, a BK accessory protein, which confers activation of BK by cGMP kinase.

KW - Antisense

KW - Guanosine 3′,5′-cyclic monophosphate

KW - Human β-subunit

KW - Large-conductance calcium-activated potassium channels

KW - Maxi-potassium

KW - Patch clamp

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M3 - Article

C2 - 12670831

AN - SCOPUS:0042570701

VL - 285

SP - F289-F294

JO - American Journal of Physiology - Renal Physiology

JF - American Journal of Physiology - Renal Physiology

SN - 0363-6127

IS - 2 54-2

ER -

Kudlacek PE, Pluznick JL, Ma R, Padanilam B, Sansom SC. Role of hβ1 in activation of human mesangial BK channels by cGMP kinase. American Journal of Physiology - Renal Physiology. 2003 Aug 1;285(2 54-2):F289-F294.