ArticleProceedings of the National Academy of Sciences of the United States of America2002
The single-channel properties of human acetylcholine alpha 7 receptors are altered by fusing alpha 7 to the green fluorescent protein.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 38 citations in OpenAlex.
- Covalent anchoring of tethered ligands to chemogenetic handles for targeted in vivo neuropharmacology.STAR protocols · 2025Article
- GFP fluorescence: A few lesser-known nuggets that make it work.Journal of biosciences · 2018Article
- Melanocortin 3 receptor has a 5' exon that directs translation of apically localized protein from the second in-frame ATG.Molecular endocrinology (Baltimore, Md.) · 2014Article
- An outline of desensitization in pentameric ligand-gated ion channel receptors.Cellular and molecular life sciences : CMLS · 2013Review
- Anterograde trafficking of neurotrophin-3 in the adult olfactory system in vivo.Experimental neurology · 2013Article
- Activation of α7 nicotinic receptors by orthosteric and allosteric agonists: influence on single-channel kinetics and conductance.Molecular pharmacology · 2012Article
- Article
- α7 nicotinic ACh receptors as a ligand-gated source of Ca(2+) ions: the search for a Ca(2+) optimum.Advances in experimental medicine and biology · 2012Review
- Trafficking of alpha4* nicotinic receptors revealed by superecliptic phluorin: effects of a beta4 amyotrophic lateral sclerosis-associated mutation and chronic exposure to nicotine.The Journal of biological chemistry · 2011Article
- Molecular dynamics and brownian dynamics investigation of ion permeation and anesthetic halothane effects on a proton-gated ion channel.Journal of the American Chemical Society · 2010Article
- Using fluorometry and ion-sensitive microelectrodes to study the functional expression of heterologously-expressed ion channels and transporters in Xenopus oocytes.Methods (San Diego, Calif.) · 2010Review
- Ric-3 chaperone-mediated stable cell-surface expression of the neuronal alpha7 nicotinic acetylcholine receptor in mammalian cells.Acta pharmacologica Sinica · 2009Article
- Green fluorescent protein: a molecular lantern that illuminates the cellular interior.Journal of biosciences · 2009Article
- Nicotinic acetylcholine receptor alpha7 subunits with a C2 cytoplasmic loop yellow fluorescent protein insertion form functional receptors.Acta pharmacologica Sinica · 2009Article
- Pentameric concatenated (alpha4)(2)(beta2)(3) and (alpha4)(3)(beta2)(2) nicotinic acetylcholine receptors: subunit arrangement determines functional expression.British journal of pharmacology · 2009Article
- A modified acetylcholine receptor delta-subunit enables a null mutant to survive beyond sexual maturation.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2008Article
- In silico models for the human alpha4beta2 nicotinic acetylcholine receptor.The journal of physical chemistry. B · 2008Article
- Properties of GluR3 receptors tagged with GFP at the amino or carboxyl terminus.Proceedings of the National Academy of Sciences of the United States of America · 2007Article
- The human adult subtype ACh receptor channel has high Ca2+ permeability and predisposes to endplate Ca2+ overloading.The Journal of physiology · 2006Article
- Membrane organization and dynamics of the G-protein-coupled serotonin1A receptor monitored using fluorescence-based approaches.Journal of fluorescence · 2005Review
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Neuronal nicotinic acetylcholine (AcCho) receptors composed of alpha7-subunits (alpha7-AcChoRs) are involved in many physiological activities. Nevertheless, very little is known about their single-channel characteristics. By using outside-out patch-clamp recordings from Xenopus oocytes expressing wild-type (wt) alpha7-AcChoRs, we identified two classes of channel conductance: a low conductance (gamma(L)) of 72 pS and a high one (gamma(H)) of 87 pS, with mean open-times (tau(op)) of 0.6 ms. The same classes of conductances, but longer tau(op) (3 ms), were seen in experiments with chimeric alpha7 receptors in which the wtalpha7 extracellular C terminus was fused to the green fluorescent protein (wtalpha7-GFP AcChoRs). In contrast, channels with three different conductances were gated by AcCho in oocytes expressing alpha7 receptors carrying a Leu-to-Thr 248 mutation (mutalpha7) or oocytes expressing chimeric mutalpha7-GFP receptors. These conductance levels were significantly smaller, and their mean open-times were larger, than those of wtalpha7-AcChoRs. Interestingly, in the absence of AcCho, these oocytes showed single-channel openings of the same conductances, but shorter tau(op), than those activated by AcCho. Accordingly, human homomeric wtalpha7 receptors open channels of high conductance and brief lifetime, and fusion to GFP lengthens their lifetime. In contrast, mutalpha7 receptors open channels of lower conductance and longer lifetime than those gated by wtalpha7-AcChoRs, and these parameters are not greatly altered by fusing the mutalpha7 to GFP. All this evidence shows that GFP-tagging can alter importantly receptor kinetics, a fact that has to be taken into account whenever tagged proteins are used to study their function.
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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.