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 cGMP signalling components such as the cGMP-regulated PDE2A (phosphodiesterase 2A) and the cGMP-gated ion channel CNGA3 (cyclic nucleotide-gated cation channel α-3). Further molecular 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.
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Research Article|
November 12 2010
Guanylate cyclase-G, expressed in the Grueneberg ganglion olfactory subsystem, is activated by bicarbonate
Ying-Chi Chao;
Ying-Chi Chao
1
*Molecular Medicine Program, Taiwan International Graduate Program, Academia Sinica, Taipei 115, Taiwan
†Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan
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Chien-Jui Cheng;
Chien-Jui Cheng
1
‡Graduate Institute of Clinical Medicine and Department of Pathology, College of Medicine, Taipei Medical University and Hospital, Taipei 110, Taiwan
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Hsiu-Ting Hsieh;
Hsiu-Ting Hsieh
†Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan
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Chih-Ching Lin;
Chih-Ching Lin
†Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan
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Chien-Chang Chen;
Chien-Chang Chen
†Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan
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Ruey-Bing Yang
Ruey-Bing Yang
2
*Molecular Medicine Program, Taiwan International Graduate Program, Academia Sinica, Taipei 115, Taiwan
†Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan
§Institute of Pharmacology, School of Medicine, National Yang-Ming University, Taipei 112, Taiwan
2To whom correspondence should be addressed (email rbyang@ibms.sinica.edu.tw).
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Publisher: Portland Press Ltd
Received:
April 22 2010
Revision Received:
July 29 2010
Accepted:
August 26 2010
Accepted Manuscript online:
August 26 2010
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2010 Biochemical Society
2010
Biochem J (2010) 432 (2): 267–273.
Article history
Received:
April 22 2010
Revision Received:
July 29 2010
Accepted:
August 26 2010
Accepted Manuscript online:
August 26 2010
Citation
Ying-Chi Chao, Chien-Jui Cheng, Hsiu-Ting Hsieh, Chih-Ching Lin, Chien-Chang Chen, Ruey-Bing Yang; Guanylate cyclase-G, expressed in the Grueneberg ganglion olfactory subsystem, is activated by bicarbonate. Biochem J 1 December 2010; 432 (2): 267–273. doi: https://doi.org/10.1042/BJ20100617
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