Subunit 2 (or β) of retinal rod cGMP-gated cation channel is a component of the 240-kDa channel-associated protein and mediates Ca2+-calmodulin modulation

Tsung-Yu Chen, M. Illing, L. L. Molday, Y. T. Hsu, K. W. Yau, R. S. Molday

Research output: Contribution to journalArticle

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Abstract

The cGMP-gated cation channel mediating visual transduction in retinal rods was recently found to comprise at least two subunits, 1 and 2 (or α and β). SDS gels of the purified channel show, in addition to a 63-kDa protein band (subunit 1), a 240-kDa protein band that binds Ca2+-calmodulin, a modulator of the channel. To examine any connection between subunit 2 and the 240-kDa protein, cGMP-gated channels formed from the expressed cloned subunits in human embryonic kidney (HEK) 293 cells were tested for Ca2+-calmodulin effect. Homooligomeric channels formed by subunit 1 alone showed no sensitivity to Ca2+-calmodulin, and neither did heterooligomeric channels formed by subunit 1 and the short alternatively spliced form of subunit 2 (2a). By contrast, the cGMP half-activation constant (K1/2) for heterooligomeric channels formed from subunit 1 and the long form of subunit 2 (2b) was increased 1.5- to 2-fold by Ca2+-calmodulin, similar to the increase observed for the native channel. In Western blots of rod outer segment membranes, a subunit 2-specific antibody also recognized the 240-kDa protein. Finally, amino acid sequences derived from peptide fragments of the bovine 240-kDa protein showed ≈80% identity to regions of subunit 2b of the human channel. These results together suggest that subunit 2b of the rod channel is a component of the 240-kDa protein and that it mediates the Ca2+-calmodulin modulation of the channel.

Original languageEnglish (US)
Pages (from-to)11757-11761
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume91
Issue number24
StatePublished - Nov 22 1994
Externally publishedYes

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Retinal Rod Photoreceptor Cells
Calmodulin
Cations
Proteins
Rod Cell Outer Segment
Peptide Fragments
Protein Subunits
Amino Acid Sequence
Western Blotting
Gels
Kidney
Membranes
Antibodies

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Subunit 2 (or β) of retinal rod cGMP-gated cation channel is a component of the 240-kDa channel-associated protein and mediates Ca2+-calmodulin modulation. / Chen, Tsung-Yu; Illing, M.; Molday, L. L.; Hsu, Y. T.; Yau, K. W.; Molday, R. S.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 91, No. 24, 22.11.1994, p. 11757-11761.

Research output: Contribution to journalArticle

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abstract = "The cGMP-gated cation channel mediating visual transduction in retinal rods was recently found to comprise at least two subunits, 1 and 2 (or α and β). SDS gels of the purified channel show, in addition to a 63-kDa protein band (subunit 1), a 240-kDa protein band that binds Ca2+-calmodulin, a modulator of the channel. To examine any connection between subunit 2 and the 240-kDa protein, cGMP-gated channels formed from the expressed cloned subunits in human embryonic kidney (HEK) 293 cells were tested for Ca2+-calmodulin effect. Homooligomeric channels formed by subunit 1 alone showed no sensitivity to Ca2+-calmodulin, and neither did heterooligomeric channels formed by subunit 1 and the short alternatively spliced form of subunit 2 (2a). By contrast, the cGMP half-activation constant (K1/2) for heterooligomeric channels formed from subunit 1 and the long form of subunit 2 (2b) was increased 1.5- to 2-fold by Ca2+-calmodulin, similar to the increase observed for the native channel. In Western blots of rod outer segment membranes, a subunit 2-specific antibody also recognized the 240-kDa protein. Finally, amino acid sequences derived from peptide fragments of the bovine 240-kDa protein showed ≈80{\%} identity to regions of subunit 2b of the human channel. These results together suggest that subunit 2b of the rod channel is a component of the 240-kDa protein and that it mediates the Ca2+-calmodulin modulation of the channel.",
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