Reconstitution and characterization of two forms of cyclic nucleotide-gated channels from skeletal muscle

Lorraine C. Santy, Guido Guidotti

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

A cyclic nucleotide-gated channel present in skeletal muscle plasma membrane has previously been identified as being responsible for insulin-activated sodium entry into muscle cells (J. E. M. McGeoch and G. Guidotti. J. Biol. Chem. 267: 832-841, 1992). We have isolated this channel activity to further study and characterize it. The channel was solubilized from rabbit skeletal muscle sarcolemma and functionally reconstituted into phospholipid vesicles, as assayed by patch-clamp analysis of the reconstituted proteins. Channel activity was isolated by 8-bromo-guanosine 3′,5′-cyclic monophosphate affinity chromatography, producing two distinct peaks of cyclic nucleotidegated channel activity. These two types of channel activity differ in guanosine 3′,5′-cyclic monophosphate affinity and in the ability to be opened by adenosine 3′,5′-cyclic monophosphate. The cyclic nucleotide-gated channel from rod outer segments also forms two peaks of activity when purified in this manner. The presence of two forms of channel activity could have implications for the mechanism of insulin-activated sodium entry.

Original languageEnglish (US)
JournalAmerican Journal of Physiology
Volume271
Issue number6 PART 1
StatePublished - Dec 1 1996

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Cyclic Nucleotide-Gated Cation Channels
Guanosine
Skeletal Muscle
Insulin
Rod Cell Outer Segment
Sarcolemma
Affinity Chromatography
Adenosine
Muscle Cells
Phospholipids
Cell Membrane
Rabbits
Proteins

All Science Journal Classification (ASJC) codes

  • Physiology (medical)

Cite this

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Reconstitution and characterization of two forms of cyclic nucleotide-gated channels from skeletal muscle. / Santy, Lorraine C.; Guidotti, Guido.

In: American Journal of Physiology, Vol. 271, No. 6 PART 1, 01.12.1996.

Research output: Contribution to journalArticle

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