Synaptic Scaffolding Molecule Binds to and Regulates Vasoactive Intestinal Polypeptide Type-1 Receptor in Epithelial Cells

Heon Yung Gee, Young Woong Kim, Min Jae Jo, Wan Namkung, Joo Young Kim, Hyun Woo Park, Kyung Sik Kim, Hoguen Kim, Akemichi Baba, Jinhee Yang, Eunjoon Kim, Kyung Hwan Kim, Min Goo Lee

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Background & Aims: Vasoactive intestinal polypeptide (VIP) is a principal regulator of fluid and electrolyte secretion in the gastrointestinal system. The VIP type-1 receptor (VPAC1), a class II G-protein-coupled receptor, contains a putative C-terminal PDZ-binding motif. A yeast 2-hybrid screen indicated that the C-terminus of VPAC1 bound to the PDZ domain of synaptic scaffolding molecule (S-SCAM, also known as membrane-associated guanylate kinase inverted-2 [MAGI-2]). We analyzed the association between S-SCAM and VPAC1. Methods: The biochemical properties and physiologic significance of the interaction between VPAC1 and S-SCAM were examined in heterologous expression systems, T84 colonic epithelial cells, and human pancreas and colon tissues using an integrated molecular and physiologic approach. Results: The physical interaction between VPAC1 and S-SCAM was confirmed by immunoprecipitation in HEK 293 mammalian cells and human pancreatic and colonic tissues. Immunocytochemical analysis indicated that S-SCAM recruited VPAC1 to the junctional area near the apical end of the lateral membrane in T84 cells. Several lines of evidence revealed that S-SCAM inhibits VPAC1 activation. Overexpression of S-SCAM inhibited VPAC1-mediated cAMP production and agonist-induced VPAC1 internalization in HEK 293 and HeLa cells. In addition, S-SCAM decreased the VPAC1-mediated current through the cystic fibrosis transmembrane conductance regulator in Xenopus oocytes, especially at low concentrations of VIP. Importantly, loss of S-SCAM increased VIP-induced short-circuit currents in T84 monolayers, which endogenously express VPAC1 and S-SCAM. Conclusions: S-SCAM/MAGI-2 interacts with and regulates VPAC1 intracellular localization in epithelial cells and inhibits VPAC1 agonist-induced activation and internalization.

Original languageEnglish
JournalGastroenterology
Volume137
Issue number2
DOIs
Publication statusPublished - 2009 Jan 1

Fingerprint

Receptors, Vasoactive Intestinal Polypeptide, Type I
Vasoactive Intestinal Peptide
Guanylate Kinases
HEK293 Cells
Epithelial Cells
PDZ Domains
Fluids and Secretions
Cystic Fibrosis Transmembrane Conductance Regulator
G-Protein-Coupled Receptors
Xenopus
HeLa Cells
Immunoprecipitation
Electrolytes
Oocytes
Pancreas
Colon
Yeasts
Membranes

All Science Journal Classification (ASJC) codes

  • Hepatology
  • Gastroenterology

Cite this

Gee, Heon Yung ; Kim, Young Woong ; Jo, Min Jae ; Namkung, Wan ; Kim, Joo Young ; Park, Hyun Woo ; Kim, Kyung Sik ; Kim, Hoguen ; Baba, Akemichi ; Yang, Jinhee ; Kim, Eunjoon ; Kim, Kyung Hwan ; Lee, Min Goo. / Synaptic Scaffolding Molecule Binds to and Regulates Vasoactive Intestinal Polypeptide Type-1 Receptor in Epithelial Cells. In: Gastroenterology. 2009 ; Vol. 137, No. 2.
@article{44e8aa27e9aa423c9252a4bbfb79b367,
title = "Synaptic Scaffolding Molecule Binds to and Regulates Vasoactive Intestinal Polypeptide Type-1 Receptor in Epithelial Cells",
abstract = "Background & Aims: Vasoactive intestinal polypeptide (VIP) is a principal regulator of fluid and electrolyte secretion in the gastrointestinal system. The VIP type-1 receptor (VPAC1), a class II G-protein-coupled receptor, contains a putative C-terminal PDZ-binding motif. A yeast 2-hybrid screen indicated that the C-terminus of VPAC1 bound to the PDZ domain of synaptic scaffolding molecule (S-SCAM, also known as membrane-associated guanylate kinase inverted-2 [MAGI-2]). We analyzed the association between S-SCAM and VPAC1. Methods: The biochemical properties and physiologic significance of the interaction between VPAC1 and S-SCAM were examined in heterologous expression systems, T84 colonic epithelial cells, and human pancreas and colon tissues using an integrated molecular and physiologic approach. Results: The physical interaction between VPAC1 and S-SCAM was confirmed by immunoprecipitation in HEK 293 mammalian cells and human pancreatic and colonic tissues. Immunocytochemical analysis indicated that S-SCAM recruited VPAC1 to the junctional area near the apical end of the lateral membrane in T84 cells. Several lines of evidence revealed that S-SCAM inhibits VPAC1 activation. Overexpression of S-SCAM inhibited VPAC1-mediated cAMP production and agonist-induced VPAC1 internalization in HEK 293 and HeLa cells. In addition, S-SCAM decreased the VPAC1-mediated current through the cystic fibrosis transmembrane conductance regulator in Xenopus oocytes, especially at low concentrations of VIP. Importantly, loss of S-SCAM increased VIP-induced short-circuit currents in T84 monolayers, which endogenously express VPAC1 and S-SCAM. Conclusions: S-SCAM/MAGI-2 interacts with and regulates VPAC1 intracellular localization in epithelial cells and inhibits VPAC1 agonist-induced activation and internalization.",
author = "Gee, {Heon Yung} and Kim, {Young Woong} and Jo, {Min Jae} and Wan Namkung and Kim, {Joo Young} and Park, {Hyun Woo} and Kim, {Kyung Sik} and Hoguen Kim and Akemichi Baba and Jinhee Yang and Eunjoon Kim and Kim, {Kyung Hwan} and Lee, {Min Goo}",
year = "2009",
month = "1",
day = "1",
doi = "10.1053/j.gastro.2009.01.065",
language = "English",
volume = "137",
journal = "Gastroenterology",
issn = "0016-5085",
publisher = "W.B. Saunders Ltd",
number = "2",

}

Synaptic Scaffolding Molecule Binds to and Regulates Vasoactive Intestinal Polypeptide Type-1 Receptor in Epithelial Cells. / Gee, Heon Yung; Kim, Young Woong; Jo, Min Jae; Namkung, Wan; Kim, Joo Young; Park, Hyun Woo; Kim, Kyung Sik; Kim, Hoguen; Baba, Akemichi; Yang, Jinhee; Kim, Eunjoon; Kim, Kyung Hwan; Lee, Min Goo.

In: Gastroenterology, Vol. 137, No. 2, 01.01.2009.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Synaptic Scaffolding Molecule Binds to and Regulates Vasoactive Intestinal Polypeptide Type-1 Receptor in Epithelial Cells

AU - Gee, Heon Yung

AU - Kim, Young Woong

AU - Jo, Min Jae

AU - Namkung, Wan

AU - Kim, Joo Young

AU - Park, Hyun Woo

AU - Kim, Kyung Sik

AU - Kim, Hoguen

AU - Baba, Akemichi

AU - Yang, Jinhee

AU - Kim, Eunjoon

AU - Kim, Kyung Hwan

AU - Lee, Min Goo

PY - 2009/1/1

Y1 - 2009/1/1

N2 - Background & Aims: Vasoactive intestinal polypeptide (VIP) is a principal regulator of fluid and electrolyte secretion in the gastrointestinal system. The VIP type-1 receptor (VPAC1), a class II G-protein-coupled receptor, contains a putative C-terminal PDZ-binding motif. A yeast 2-hybrid screen indicated that the C-terminus of VPAC1 bound to the PDZ domain of synaptic scaffolding molecule (S-SCAM, also known as membrane-associated guanylate kinase inverted-2 [MAGI-2]). We analyzed the association between S-SCAM and VPAC1. Methods: The biochemical properties and physiologic significance of the interaction between VPAC1 and S-SCAM were examined in heterologous expression systems, T84 colonic epithelial cells, and human pancreas and colon tissues using an integrated molecular and physiologic approach. Results: The physical interaction between VPAC1 and S-SCAM was confirmed by immunoprecipitation in HEK 293 mammalian cells and human pancreatic and colonic tissues. Immunocytochemical analysis indicated that S-SCAM recruited VPAC1 to the junctional area near the apical end of the lateral membrane in T84 cells. Several lines of evidence revealed that S-SCAM inhibits VPAC1 activation. Overexpression of S-SCAM inhibited VPAC1-mediated cAMP production and agonist-induced VPAC1 internalization in HEK 293 and HeLa cells. In addition, S-SCAM decreased the VPAC1-mediated current through the cystic fibrosis transmembrane conductance regulator in Xenopus oocytes, especially at low concentrations of VIP. Importantly, loss of S-SCAM increased VIP-induced short-circuit currents in T84 monolayers, which endogenously express VPAC1 and S-SCAM. Conclusions: S-SCAM/MAGI-2 interacts with and regulates VPAC1 intracellular localization in epithelial cells and inhibits VPAC1 agonist-induced activation and internalization.

AB - Background & Aims: Vasoactive intestinal polypeptide (VIP) is a principal regulator of fluid and electrolyte secretion in the gastrointestinal system. The VIP type-1 receptor (VPAC1), a class II G-protein-coupled receptor, contains a putative C-terminal PDZ-binding motif. A yeast 2-hybrid screen indicated that the C-terminus of VPAC1 bound to the PDZ domain of synaptic scaffolding molecule (S-SCAM, also known as membrane-associated guanylate kinase inverted-2 [MAGI-2]). We analyzed the association between S-SCAM and VPAC1. Methods: The biochemical properties and physiologic significance of the interaction between VPAC1 and S-SCAM were examined in heterologous expression systems, T84 colonic epithelial cells, and human pancreas and colon tissues using an integrated molecular and physiologic approach. Results: The physical interaction between VPAC1 and S-SCAM was confirmed by immunoprecipitation in HEK 293 mammalian cells and human pancreatic and colonic tissues. Immunocytochemical analysis indicated that S-SCAM recruited VPAC1 to the junctional area near the apical end of the lateral membrane in T84 cells. Several lines of evidence revealed that S-SCAM inhibits VPAC1 activation. Overexpression of S-SCAM inhibited VPAC1-mediated cAMP production and agonist-induced VPAC1 internalization in HEK 293 and HeLa cells. In addition, S-SCAM decreased the VPAC1-mediated current through the cystic fibrosis transmembrane conductance regulator in Xenopus oocytes, especially at low concentrations of VIP. Importantly, loss of S-SCAM increased VIP-induced short-circuit currents in T84 monolayers, which endogenously express VPAC1 and S-SCAM. Conclusions: S-SCAM/MAGI-2 interacts with and regulates VPAC1 intracellular localization in epithelial cells and inhibits VPAC1 agonist-induced activation and internalization.

UR - http://www.scopus.com/inward/record.url?scp=67650951122&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67650951122&partnerID=8YFLogxK

U2 - 10.1053/j.gastro.2009.01.065

DO - 10.1053/j.gastro.2009.01.065

M3 - Article

VL - 137

JO - Gastroenterology

JF - Gastroenterology

SN - 0016-5085

IS - 2

ER -