Isolation and characterization of the α-subunit of the rat rod photoreceptor cGMP-gated cation channel

Colin Barnstable, Ji Ye Wei

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

A combination of genomic and PCR clones has been used to derive the full-length coding sequence of the α-subunit of the rat rod photoreceptor cGMP-gated cation channel. The sequence encodes a protein of 683 amino acids with a predicted molecular weight of 79,221. The sequence shows extensive homology with other rod cGMP-gated channels and also with the rat olfactory cyclic nucleotide-gated cation channel. When the full-length sequence of the rat rod channel was expressed in Xenopus oocytes it gave a conductance that responded to cGMP with an EC50 of 42 μM. No response to 2 m M cAMP was detected. The conductance was decreased in the presence of increasing concentrations of calcium. Both monoclonal and polyclonal antibodies were generated against a C-terminal peptide of the rat rod channel. On immunoblots of adult rat retinal membranes the antibodies recognized a band of 71 kDa, suggesting that the rat channel may undergo proteolytic cleavage in the retina, as has previously been found for the bovine rod channel. Immunocytochemical labeling of adult rat retinal sections detected prominent labeling over the rod photoreceptor outer segments with both monoclonal and polyclonal antibodies.

Original languageEnglish (US)
Pages (from-to)289-302
Number of pages14
JournalJournal of Molecular Neuroscience
Volume6
Issue number4
DOIs
StatePublished - Dec 1 1995

Fingerprint

Retinal Rod Photoreceptor Cells
Cations
Monoclonal Antibodies
Rod Cell Outer Segment
Xenopus
Oocytes
Retina
Clone Cells
Molecular Weight
Calcium
Amino Acids
Polymerase Chain Reaction
Peptides
Membranes
Antibodies
Proteins

All Science Journal Classification (ASJC) codes

  • Cellular and Molecular Neuroscience

Cite this

@article{3f72f18c3ba14fa0843994d207785e33,
title = "Isolation and characterization of the α-subunit of the rat rod photoreceptor cGMP-gated cation channel",
abstract = "A combination of genomic and PCR clones has been used to derive the full-length coding sequence of the α-subunit of the rat rod photoreceptor cGMP-gated cation channel. The sequence encodes a protein of 683 amino acids with a predicted molecular weight of 79,221. The sequence shows extensive homology with other rod cGMP-gated channels and also with the rat olfactory cyclic nucleotide-gated cation channel. When the full-length sequence of the rat rod channel was expressed in Xenopus oocytes it gave a conductance that responded to cGMP with an EC50 of 42 μM. No response to 2 m M cAMP was detected. The conductance was decreased in the presence of increasing concentrations of calcium. Both monoclonal and polyclonal antibodies were generated against a C-terminal peptide of the rat rod channel. On immunoblots of adult rat retinal membranes the antibodies recognized a band of 71 kDa, suggesting that the rat channel may undergo proteolytic cleavage in the retina, as has previously been found for the bovine rod channel. Immunocytochemical labeling of adult rat retinal sections detected prominent labeling over the rod photoreceptor outer segments with both monoclonal and polyclonal antibodies.",
author = "Colin Barnstable and Wei, {Ji Ye}",
year = "1995",
month = "12",
day = "1",
doi = "10.1007/BF02736787",
language = "English (US)",
volume = "6",
pages = "289--302",
journal = "Journal of Molecular Neuroscience",
issn = "0895-8696",
publisher = "Humana Press",
number = "4",

}

Isolation and characterization of the α-subunit of the rat rod photoreceptor cGMP-gated cation channel. / Barnstable, Colin; Wei, Ji Ye.

In: Journal of Molecular Neuroscience, Vol. 6, No. 4, 01.12.1995, p. 289-302.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Isolation and characterization of the α-subunit of the rat rod photoreceptor cGMP-gated cation channel

AU - Barnstable, Colin

AU - Wei, Ji Ye

PY - 1995/12/1

Y1 - 1995/12/1

N2 - A combination of genomic and PCR clones has been used to derive the full-length coding sequence of the α-subunit of the rat rod photoreceptor cGMP-gated cation channel. The sequence encodes a protein of 683 amino acids with a predicted molecular weight of 79,221. The sequence shows extensive homology with other rod cGMP-gated channels and also with the rat olfactory cyclic nucleotide-gated cation channel. When the full-length sequence of the rat rod channel was expressed in Xenopus oocytes it gave a conductance that responded to cGMP with an EC50 of 42 μM. No response to 2 m M cAMP was detected. The conductance was decreased in the presence of increasing concentrations of calcium. Both monoclonal and polyclonal antibodies were generated against a C-terminal peptide of the rat rod channel. On immunoblots of adult rat retinal membranes the antibodies recognized a band of 71 kDa, suggesting that the rat channel may undergo proteolytic cleavage in the retina, as has previously been found for the bovine rod channel. Immunocytochemical labeling of adult rat retinal sections detected prominent labeling over the rod photoreceptor outer segments with both monoclonal and polyclonal antibodies.

AB - A combination of genomic and PCR clones has been used to derive the full-length coding sequence of the α-subunit of the rat rod photoreceptor cGMP-gated cation channel. The sequence encodes a protein of 683 amino acids with a predicted molecular weight of 79,221. The sequence shows extensive homology with other rod cGMP-gated channels and also with the rat olfactory cyclic nucleotide-gated cation channel. When the full-length sequence of the rat rod channel was expressed in Xenopus oocytes it gave a conductance that responded to cGMP with an EC50 of 42 μM. No response to 2 m M cAMP was detected. The conductance was decreased in the presence of increasing concentrations of calcium. Both monoclonal and polyclonal antibodies were generated against a C-terminal peptide of the rat rod channel. On immunoblots of adult rat retinal membranes the antibodies recognized a band of 71 kDa, suggesting that the rat channel may undergo proteolytic cleavage in the retina, as has previously been found for the bovine rod channel. Immunocytochemical labeling of adult rat retinal sections detected prominent labeling over the rod photoreceptor outer segments with both monoclonal and polyclonal antibodies.

UR - http://www.scopus.com/inward/record.url?scp=0029421628&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029421628&partnerID=8YFLogxK

U2 - 10.1007/BF02736787

DO - 10.1007/BF02736787

M3 - Article

VL - 6

SP - 289

EP - 302

JO - Journal of Molecular Neuroscience

JF - Journal of Molecular Neuroscience

SN - 0895-8696

IS - 4

ER -