Angiotensin II inhibits activity of human organic anion transporter 3 through activation of protein kinase Cα: Accelerating endocytosis of the transporter

Peng Duan, Shanshan Li, Guofeng You

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Human organic anion transporter 3 (hOAT3) belongs to a family of organic anion transporters that play critical roles in the body disposition of numerous clinically important drugs. In the kidney, hOAT3 functions through a tertiary transport mechanism involving two other membrane proteins Na/K-ATPase and Na-dicarboxylate cotransporter. In the current study, we established COS-7 cells stably expressing hOAT3 and examined the regulation of hOAT3 by protein kinase C (PKC) and angiotensin II. Both PKC activation and angiotensin II inhibited hOAT3 transport activity. Angiotensin II induced inhibition of hOAT3 activity could be prevented by treating hOAT3-expressing cells with staurosporine, a general inhibitor for PKC, and with Gö6976 (5,6,7,13-Tetrahydro-13-methyl-5-oxo-12H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-12-propanenitrile), a PKCα-specific inhibitor. Examination of hOAT3 expression and transport kinetics revealed that angiotensin II induced inhibition of hOAT3 activity mainly resulted from a decreased cell surface expression kinetically reflected as a decreased Vmax without a significant change in Km. Such angiotensin II induced decrease in cell surface expression of hOAT3 was caused by an increase in hOAT3 endocytosis. However, angiotensin II induced endocytosis of Na/K-ATPase did not occur under such condition. We concluded that angiotensin II inhibited hOAT3 activity through the activation of PKCα, which led to an acceleration of hOAT3 endocytosis.

Original languageEnglish (US)
Pages (from-to)49-55
Number of pages7
JournalEuropean Journal of Pharmacology
Volume627
Issue number1-3
DOIs
StatePublished - Feb 10 2010

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Organic Anion Transporters
Endocytosis
Human Activities
Angiotensin II
Protein Kinase C
Staurosporine
COS Cells

All Science Journal Classification (ASJC) codes

  • Pharmacology

Keywords

  • Angiotensin II
  • COS-7 Cells
  • Drug transporter
  • Endocytosis
  • Protein Kinase C
  • Regulation

Cite this

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title = "Angiotensin II inhibits activity of human organic anion transporter 3 through activation of protein kinase Cα: Accelerating endocytosis of the transporter",
abstract = "Human organic anion transporter 3 (hOAT3) belongs to a family of organic anion transporters that play critical roles in the body disposition of numerous clinically important drugs. In the kidney, hOAT3 functions through a tertiary transport mechanism involving two other membrane proteins Na/K-ATPase and Na-dicarboxylate cotransporter. In the current study, we established COS-7 cells stably expressing hOAT3 and examined the regulation of hOAT3 by protein kinase C (PKC) and angiotensin II. Both PKC activation and angiotensin II inhibited hOAT3 transport activity. Angiotensin II induced inhibition of hOAT3 activity could be prevented by treating hOAT3-expressing cells with staurosporine, a general inhibitor for PKC, and with G{\"o}6976 (5,6,7,13-Tetrahydro-13-methyl-5-oxo-12H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-12-propanenitrile), a PKCα-specific inhibitor. Examination of hOAT3 expression and transport kinetics revealed that angiotensin II induced inhibition of hOAT3 activity mainly resulted from a decreased cell surface expression kinetically reflected as a decreased Vmax without a significant change in Km. Such angiotensin II induced decrease in cell surface expression of hOAT3 was caused by an increase in hOAT3 endocytosis. However, angiotensin II induced endocytosis of Na/K-ATPase did not occur under such condition. We concluded that angiotensin II inhibited hOAT3 activity through the activation of PKCα, which led to an acceleration of hOAT3 endocytosis.",
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author = "Peng Duan and Shanshan Li and Guofeng You",
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T1 - Angiotensin II inhibits activity of human organic anion transporter 3 through activation of protein kinase Cα

T2 - Accelerating endocytosis of the transporter

AU - Duan, Peng

AU - Li, Shanshan

AU - You, Guofeng

PY - 2010/2/10

Y1 - 2010/2/10

N2 - Human organic anion transporter 3 (hOAT3) belongs to a family of organic anion transporters that play critical roles in the body disposition of numerous clinically important drugs. In the kidney, hOAT3 functions through a tertiary transport mechanism involving two other membrane proteins Na/K-ATPase and Na-dicarboxylate cotransporter. In the current study, we established COS-7 cells stably expressing hOAT3 and examined the regulation of hOAT3 by protein kinase C (PKC) and angiotensin II. Both PKC activation and angiotensin II inhibited hOAT3 transport activity. Angiotensin II induced inhibition of hOAT3 activity could be prevented by treating hOAT3-expressing cells with staurosporine, a general inhibitor for PKC, and with Gö6976 (5,6,7,13-Tetrahydro-13-methyl-5-oxo-12H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-12-propanenitrile), a PKCα-specific inhibitor. Examination of hOAT3 expression and transport kinetics revealed that angiotensin II induced inhibition of hOAT3 activity mainly resulted from a decreased cell surface expression kinetically reflected as a decreased Vmax without a significant change in Km. Such angiotensin II induced decrease in cell surface expression of hOAT3 was caused by an increase in hOAT3 endocytosis. However, angiotensin II induced endocytosis of Na/K-ATPase did not occur under such condition. We concluded that angiotensin II inhibited hOAT3 activity through the activation of PKCα, which led to an acceleration of hOAT3 endocytosis.

AB - Human organic anion transporter 3 (hOAT3) belongs to a family of organic anion transporters that play critical roles in the body disposition of numerous clinically important drugs. In the kidney, hOAT3 functions through a tertiary transport mechanism involving two other membrane proteins Na/K-ATPase and Na-dicarboxylate cotransporter. In the current study, we established COS-7 cells stably expressing hOAT3 and examined the regulation of hOAT3 by protein kinase C (PKC) and angiotensin II. Both PKC activation and angiotensin II inhibited hOAT3 transport activity. Angiotensin II induced inhibition of hOAT3 activity could be prevented by treating hOAT3-expressing cells with staurosporine, a general inhibitor for PKC, and with Gö6976 (5,6,7,13-Tetrahydro-13-methyl-5-oxo-12H-indolo[2,3-a]pyrrolo[3,4-c]carbazole-12-propanenitrile), a PKCα-specific inhibitor. Examination of hOAT3 expression and transport kinetics revealed that angiotensin II induced inhibition of hOAT3 activity mainly resulted from a decreased cell surface expression kinetically reflected as a decreased Vmax without a significant change in Km. Such angiotensin II induced decrease in cell surface expression of hOAT3 was caused by an increase in hOAT3 endocytosis. However, angiotensin II induced endocytosis of Na/K-ATPase did not occur under such condition. We concluded that angiotensin II inhibited hOAT3 activity through the activation of PKCα, which led to an acceleration of hOAT3 endocytosis.

KW - Angiotensin II

KW - COS-7 Cells

KW - Drug transporter

KW - Endocytosis

KW - Protein Kinase C

KW - Regulation

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