Guanylate cyclase-G, expressed in the Grueneberg ganglion olfactory subsystem, is activated by bicarbonate

Ying Chi Chao, Chien Jui Cheng, Hsiu Ting Hsieh, Chih Ching Lin, Chien Chang Chen, Ruey Bing Yang

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

GC (guanylate cyclase)-G is the most recently identified member of the receptor GC family. However, the regulation of its activity and protein expression in the mammalian olfactory system remains unclear. In the present study, we used a GC-G-specific antibody to validate that the GC-G protein is expressed in Grueneberg ganglion neurons, a newly recognized olfactory subsystem co-expressing other cGMPsignalling components such as the cGMP-regulated PDE2A (phosphodiesterase 2A) and the cGMP-gated ion channel CNGA3 (cyclic nucleotide-gated cation channel α-3). Furthermolecular and biochemical analyses showed that heterologously expressed GC-G protein, specifically the C-terminal cyclase domain, was directly stimulated by bicarbonate in both in vivo cellular cGMP accumulation assays in human embryonic kidney-293T cells and in vitro GC assays with a purified recombinant protein containing the GC domain. In addition, overexpression of GC-G in NG108 neuronal cells resulted in a CO2-dependent increase in cellular cGMP level that could be blocked by treatment with acetazolamide, an inhibitor of carbonic anhydrases, which implies that the stimulatory effect of CO2 requires its conversion to bicarbonate. Together, our data demonstrate a novel CO2/bicarbonate- dependent activation mechanism for GC-G and suggest that GC-G may be involved in a wide variety of CO2/bicarbonate-regulated biological processes such as the chemosensory function in Grueneberg ganglion neurons.

Original languageEnglish
Pages (from-to)267-273
Number of pages7
JournalBiochemical Journal
Volume432
Issue number2
DOIs
Publication statusPublished - Dec 1 2010

Fingerprint

Guanylate Cyclase
Bicarbonates
Ganglia
GTP-Binding Proteins
Neurons
Type 2 Cyclic Nucleotide Phosphodiesterases
Assays
Cyclic Nucleotide-Gated Cation Channels
Carbonic Anhydrase Inhibitors
Biological Phenomena
Acetazolamide
HEK293 Cells
Protein C
Recombinant Proteins
Chemical activation
Kidney
Antibodies

Keywords

  • Bicarbonate
  • cGMP
  • Grueneburg ganglion (GG)
  • Guanylate cyclase (GC)
  • Receptor
  • Signal transduction

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Guanylate cyclase-G, expressed in the Grueneberg ganglion olfactory subsystem, is activated by bicarbonate. / Chao, Ying Chi; Cheng, Chien Jui; Hsieh, Hsiu Ting; Lin, Chih Ching; Chen, Chien Chang; Yang, Ruey Bing.

In: Biochemical Journal, Vol. 432, No. 2, 01.12.2010, p. 267-273.

Research output: Contribution to journalArticle

Chao, Ying Chi ; Cheng, Chien Jui ; Hsieh, Hsiu Ting ; Lin, Chih Ching ; Chen, Chien Chang ; Yang, Ruey Bing. / Guanylate cyclase-G, expressed in the Grueneberg ganglion olfactory subsystem, is activated by bicarbonate. In: Biochemical Journal. 2010 ; Vol. 432, No. 2. pp. 267-273.
@article{1f08449b6dac46b98a8c7863806dd6db,
title = "Guanylate cyclase-G, expressed in the Grueneberg ganglion olfactory subsystem, is activated by bicarbonate",
abstract = "GC (guanylate cyclase)-G is the most recently identified member of the receptor GC family. However, the regulation of its activity and protein expression in the mammalian olfactory system remains unclear. In the present study, we used a GC-G-specific antibody to validate that the GC-G protein is expressed in Grueneberg ganglion neurons, a newly recognized olfactory subsystem co-expressing other cGMPsignalling components such as the cGMP-regulated PDE2A (phosphodiesterase 2A) and the cGMP-gated ion channel CNGA3 (cyclic nucleotide-gated cation channel α-3). Furthermolecular and biochemical analyses showed that heterologously expressed GC-G protein, specifically the C-terminal cyclase domain, was directly stimulated by bicarbonate in both in vivo cellular cGMP accumulation assays in human embryonic kidney-293T cells and in vitro GC assays with a purified recombinant protein containing the GC domain. In addition, overexpression of GC-G in NG108 neuronal cells resulted in a CO2-dependent increase in cellular cGMP level that could be blocked by treatment with acetazolamide, an inhibitor of carbonic anhydrases, which implies that the stimulatory effect of CO2 requires its conversion to bicarbonate. Together, our data demonstrate a novel CO2/bicarbonate- dependent activation mechanism for GC-G and suggest that GC-G may be involved in a wide variety of CO2/bicarbonate-regulated biological processes such as the chemosensory function in Grueneberg ganglion neurons.",
keywords = "Bicarbonate, cGMP, Grueneburg ganglion (GG), Guanylate cyclase (GC), Receptor, Signal transduction",
author = "Chao, {Ying Chi} and Cheng, {Chien Jui} and Hsieh, {Hsiu Ting} and Lin, {Chih Ching} and Chen, {Chien Chang} and Yang, {Ruey Bing}",
year = "2010",
month = "12",
day = "1",
doi = "10.1042/BJ20100617",
language = "English",
volume = "432",
pages = "267--273",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "2",

}

TY - JOUR

T1 - Guanylate cyclase-G, expressed in the Grueneberg ganglion olfactory subsystem, is activated by bicarbonate

AU - Chao, Ying Chi

AU - Cheng, Chien Jui

AU - Hsieh, Hsiu Ting

AU - Lin, Chih Ching

AU - Chen, Chien Chang

AU - Yang, Ruey Bing

PY - 2010/12/1

Y1 - 2010/12/1

N2 - GC (guanylate cyclase)-G is the most recently identified member of the receptor GC family. However, the regulation of its activity and protein expression in the mammalian olfactory system remains unclear. In the present study, we used a GC-G-specific antibody to validate that the GC-G protein is expressed in Grueneberg ganglion neurons, a newly recognized olfactory subsystem co-expressing other cGMPsignalling components such as the cGMP-regulated PDE2A (phosphodiesterase 2A) and the cGMP-gated ion channel CNGA3 (cyclic nucleotide-gated cation channel α-3). Furthermolecular and biochemical analyses showed that heterologously expressed GC-G protein, specifically the C-terminal cyclase domain, was directly stimulated by bicarbonate in both in vivo cellular cGMP accumulation assays in human embryonic kidney-293T cells and in vitro GC assays with a purified recombinant protein containing the GC domain. In addition, overexpression of GC-G in NG108 neuronal cells resulted in a CO2-dependent increase in cellular cGMP level that could be blocked by treatment with acetazolamide, an inhibitor of carbonic anhydrases, which implies that the stimulatory effect of CO2 requires its conversion to bicarbonate. Together, our data demonstrate a novel CO2/bicarbonate- dependent activation mechanism for GC-G and suggest that GC-G may be involved in a wide variety of CO2/bicarbonate-regulated biological processes such as the chemosensory function in Grueneberg ganglion neurons.

AB - GC (guanylate cyclase)-G is the most recently identified member of the receptor GC family. However, the regulation of its activity and protein expression in the mammalian olfactory system remains unclear. In the present study, we used a GC-G-specific antibody to validate that the GC-G protein is expressed in Grueneberg ganglion neurons, a newly recognized olfactory subsystem co-expressing other cGMPsignalling components such as the cGMP-regulated PDE2A (phosphodiesterase 2A) and the cGMP-gated ion channel CNGA3 (cyclic nucleotide-gated cation channel α-3). Furthermolecular and biochemical analyses showed that heterologously expressed GC-G protein, specifically the C-terminal cyclase domain, was directly stimulated by bicarbonate in both in vivo cellular cGMP accumulation assays in human embryonic kidney-293T cells and in vitro GC assays with a purified recombinant protein containing the GC domain. In addition, overexpression of GC-G in NG108 neuronal cells resulted in a CO2-dependent increase in cellular cGMP level that could be blocked by treatment with acetazolamide, an inhibitor of carbonic anhydrases, which implies that the stimulatory effect of CO2 requires its conversion to bicarbonate. Together, our data demonstrate a novel CO2/bicarbonate- dependent activation mechanism for GC-G and suggest that GC-G may be involved in a wide variety of CO2/bicarbonate-regulated biological processes such as the chemosensory function in Grueneberg ganglion neurons.

KW - Bicarbonate

KW - cGMP

KW - Grueneburg ganglion (GG)

KW - Guanylate cyclase (GC)

KW - Receptor

KW - Signal transduction

UR - http://www.scopus.com/inward/record.url?scp=78649590775&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78649590775&partnerID=8YFLogxK

U2 - 10.1042/BJ20100617

DO - 10.1042/BJ20100617

M3 - Article

C2 - 20738256

AN - SCOPUS:78649590775

VL - 432

SP - 267

EP - 273

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 2

ER -