Evidence map›Paper›PMID 21364764›Full record

ArticlePloS one2011

The roles of transmembrane domain helix-III during rhodopsin photoactivation.

Wen-bin Ou, Tingfang Yi, Jong-Myoung Kim, H Gobind Khorana

Open access · goldAbstract read
In one paragraph

Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.

0numbers the graph read from it
0cells of the map it votes in
5citing papers in PubMed
1.1field-weighted citation impact, top 24% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

5 citing papers in PubMed, 12 citations in OpenAlex.

  1. Article
  2. Article
  3. HeterozygousBioscience reports · 2019
    Article
  4. Article
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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

4 authors at 2 institutions in 1 country.

Wen-bin OuDepartment of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America. ouwb75@gmail.com
Tingfang Yi
Jong-Myoung Kim
H Gobind Khorana
Massachusetts Institute of Technology · USHarvard University · US

Funding

SYNTHETIC AND BIOLOGICAL STUDIES OF NUCLEIC ACIDSR01GM028289 · NIGMS · MASSACHUSETTS INSTITUTE OF TECHNOLOGY · PI KHORANA, H. GOBIND · 1985 to 2003
$2.9M
RHODOPSIN MISFOLDING IN RETINITIS PIGMENTOSAR01EY011716 · NEI · MASSACHUSETTS INSTITUTE OF TECHNOLOGY · PI KHORANA, H. GOBIND · 1998 to 2007
$1.7M
SYNTHETIC &BIOLOGICAL STUDIES OF NUCLEIC ACIDSR37GM028289 · NIGMS · MASSACHUSETTS INSTITUTE OF TECHNOLOGY · PI KHORANA, H. GOBIND · 1986 to 1995
–
NEI NIH HHS EY11716NEI NIH HHS R01 EY011716NIGMS NIH HHS GM28289NIGMS NIH HHS R01 GM028289
6 · The paper itself

Abstract

backgroundRhodopsin, the prototypic member of G protein-coupled receptors (GPCRs), undergoes isomerization of 11-cis-retinal to all-trans-retinal upon photoactivation. Although the basic mechanism by which rhodopsin is activated is well understood, the roles of whole transmembrane (TM) helix-III during rhodopsin photoactivation in detail are not completely clear. PRINCIPAL

findingsWe herein use single-cysteine mutagenesis technique to investigate conformational changes in TM helices of rhodopsin upon photoactivation. Specifically, we study changes in accessibility and reactivity of cysteine residues introduced into the TM helix-III of rhodopsin. Twenty-eight single-cysteine mutants of rhodopsin (P107C-R135C) were prepared after substitution of all natural cysteine residues (C140/C167/C185/C222/C264/C316) by alanine. The cysteine mutants were expressed in COS-1 cells and rhodopsin was purified after regeneration with 11-cis-retinal. Cysteine accessibility in these mutants was monitored by reaction with 4, 4'-dithiodipyridine (4-PDS) in the dark and after illumination. Most of the mutants except for T108C, G109C, E113C, I133C, and R135C showed no reaction in the dark. Wide variation in reactivity was observed among cysteines at different positions in the sequence 108-135 after photoactivation. In particular, cysteines at position 115, 119, 121, 129, 131, 132, and 135, facing 11-cis-retinal, reacted with 4-PDS faster than neighboring amino acids. The different reaction rates of mutants with 4-PDS after photoactivation suggest that the amino acids in different positions in helix-III are exposed to aqueous environment to varying degrees. SIGNIFICANCE: Accessibility data indicate that an aqueous/hydrophobic boundary in helix-III is near G109 and I133. The lack of reactivity in the dark and the accessibility of cysteine after photoactivation indicate an increase of water/4-PDS accessibility for certain cysteine-mutants at Helix-III during formation of Meta II. We conclude that photoactivation resulted in water-accessible at the chromophore-facing residues of Helix-III.

Indexed as

Photochemical ProcessesAmino Acid SequenceAnimalsCatalysisChlorocebus aethiopsCOS CellsCysteineLightMembrane ProteinsModels, BiologicalMolecular Sequence DataMutagenesis, Site-DirectedMutant ProteinsProtein Interaction Domains and MotifsProtein Structure, SecondaryRhodopsinCysteineMembrane ProteinsMutant ProteinsRhodopsin

Identifiers

PMID21364764
PMCPMC3045455
OpenAlexW2056936342

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.