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PMID: 11842144 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Electrophysiological analysis of cloned cyclic nucleotide-gated ion channels.

Plant physiology ·Vol. 128 ·No. 2 ·2002-02-00 ·Pages 400-10

Leng Q, Mercier RW, Hua BG, Fromm H, Berkowitz GA

Abstract

Electrophysiological studies were conducted on the cloned plant cyclic nucleotide-gated ion channels AtCNGC2 and AtCNGC1 from Arabidopsis, and NtCBP4 from tobacco (Nicotiana tobacum). The nucleotide coding sequences for these proteins were expressed in Xenopus laevis oocytes or HEK 293 cells. Channel characteristics were evaluated using voltage clamp analysis of currents in the presence of cAMP. AtCNGC2 was demonstrated to conduct K(+) and other monovalent cations, but exclude Na(+); this conductivity profile is unique for any ion channel not possessing the amino acid sequence found in the selectivity filter of K(+)-selective ion channels. Application of cAMP evoked currents in membrane patches of oocytes injected with AtCNGC2 cRNA. Direct activation of the channel by cyclic nucleotide, demonstrated by application of cyclic nucleotide to patches of membranes expressing such channels, is a hallmark characteristic of this ion channel family. Voltage clamp studies (two-electrode configuration) demonstrated that AtCNGC1 and NtCBP4 are also cyclic nucleotide-gated channels. Addition of a lipophilic analog of cAMP to the perfusion bath of oocytes injected with NtCBP4 and AtCNGC1 cRNAs induced inward rectified, noninactivating K(+) currents.

MeSH Terms
Amino Acid Sequence Animals Arabidopsis/genetics,physiology Calmodulin-Binding Proteins/genetics,metabolism Cell Line Cloning, Molecular Cyclic AMP/pharmacology Cyclic Nucleotide-Gated Cation Channels Gene Expression Humans Ion Channels/genetics,physiology Ion Transport/drug effects,physiology Ligands Membrane Potentials/drug effects,physiology Molecular Sequence Data Oocytes/physiology Patch-Clamp Techniques Plant Proteins/genetics,physiology Potassium/pharmacology RNA, Complementary/genetics Sequence Homology, Amino Acid Sodium/pharmacology Tobacco/genetics,physiology Xenopus laevis
Chemicals
CBP4 protein, Nicotiana tabacum Calmodulin-Binding Proteins Cyclic Nucleotide-Gated Cation Channels Ion Channels Ligands Plant Proteins RNA, Complementary Sodium Cyclic AMP Potassium
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Leng Qiang
Department of Plant Science, University of Connecticut, Storrs, CT 06269-4067, USA.
Mercier Richard W
Hua Bao-Guang
Fromm Hillel
Berkowitz Gerald A
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Article Info
Journal
Plant physiology
Abbr.
Plant Physiol
ISSN
0032-0889
DOI
10.1104/pp.010832
Published
2002-02-00
Pages
400-10
Language
English
Region
United States
NLM ID
0401224
PMCID
PMC148903
Subset
IM
External Links
PubMed source DOI: 10.1104/pp.010832 PMC Full Text View journal details
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