A novel α-conotoxin, PeIA, cloned from Conus pergrandis, discriminates between Rat α9α10 and α7 nicotinic cholinergic receptors
The α9 and α10 nicotinic cholinergic subunits assemble to form the receptor believed to mediate synaptic transmission between efferent olivocochlear fibers and hair cells of the cochlea, one of the few examples of postsynaptic function for a non-muscle nicotinic acetylcholine receptor (nAChR). Howev...
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2005
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v280_n34_p30107_McIntosh http://hdl.handle.net/20.500.12110/paper_00219258_v280_n34_p30107_McIntosh |
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paper:paper_00219258_v280_n34_p30107_McIntosh2023-06-08T14:43:26Z A novel α-conotoxin, PeIA, cloned from Conus pergrandis, discriminates between Rat α9α10 and α7 nicotinic cholinergic receptors Biosynthesis Blood Cells Characterization Cloning Molecular biology Muscle Skin Cochlea Hair cells Thymocytes Xenopus oocytes Biochemistry acetylcholine alpha bungarotoxin alpha conotoxin PeIA bungarotoxin receptor chimeric protein conotoxin nicotinic alpha9alpha10 receptor nicotinic receptor protein subunit receptor subtype recombinant protein snail venom unclassified drug animal cell article binding affinity cholinergic receptor blocking cochlea cochlear nerve concentration response controlled study Conus pergrandis DNA sequence genetic variability hair cell IC 50 matrix assisted laser desorption ionization time of flight mass spectrometry molecular cloning molecular evolution nonhuman oocyte priority journal protein determination protein expression protein function protein localization rat receptor binding receptor sensitivity sequence analysis snail synaptic transmission toxin analysis toxin synthesis wild type Xenopus Acetylcholine Amino Acid Sequence Animals Base Sequence Bungarotoxins Cell Differentiation Cloning, Molecular Conotoxins Dose-Response Relationship, Drug Electrophysiology Ganglia, Spinal Hair Cells, Inner Inhibitory Concentration 50 Kinetics Lymphocytes Molecular Sequence Data Mollusca Neurons Oocytes Peptides Protein Subunits Rats Rats, Sprague-Dawley Receptors, Nicotinic Recombinant Proteins RNA Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Synaptic Transmission Thymus Gland Xenopus Xenopus laevis Conus geographus Conus magus The α9 and α10 nicotinic cholinergic subunits assemble to form the receptor believed to mediate synaptic transmission between efferent olivocochlear fibers and hair cells of the cochlea, one of the few examples of postsynaptic function for a non-muscle nicotinic acetylcholine receptor (nAChR). However, it has been suggested that the expression profile of α9 and α10 overlaps with that of α7 in the cochlea and in sites such as dorsal root ganglion neurons, peripheral blood lymphocytes, developing thymocytes, and skin. We now report the cloning, total synthesis, and characterization of a novel toxin α-conotoxin PeIA that discriminates between α9α10 and α7 nAChRs. This is the first toxin to be identified from Conus pergrandis, a species found in deep waters of the Western Pacific. α-Conotoxin PeIA displayed a 260-fold higher selectivity for α-bungarotoxin-sensitive α9α10 nAChRs compared with α-bungarotoxin-sensitive α7 receptors. The IC50 of the toxin was 6.9 ± 0.5 nM and 4.4 ± 0.5 nM for recombinant α9α10 and wild-type hair cell nAChRs, respectively. α-Conotoxin PeIA bears high resemblance to α-conotoxins MII and GIC isolated from Conus magus and Conus geographus, respectively. However, neither α-conotoxin MII nor α-conotoxin GIC at concentrations of 10 μM blocked acetylcholine responses elicited in Xenopus oocytes injected with the α9 and α10 subunits. Among neuronal non-α-bungarotoxin- sensitive receptors, α-conotoxin PeIA was also active at α3β2 receptors and chimeric α6/α3β2β3 receptors. α-Conotoxin PeIA represents a novel probe to differentiate responses mediated either through α9α10 or α7 nAChRs in those tissues where both receptors are expressed. © 2005 by The American Society for Biochemistry and Molecular Biology, Inc. 2005 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v280_n34_p30107_McIntosh http://hdl.handle.net/20.500.12110/paper_00219258_v280_n34_p30107_McIntosh |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Biosynthesis Blood Cells Characterization Cloning Molecular biology Muscle Skin Cochlea Hair cells Thymocytes Xenopus oocytes Biochemistry acetylcholine alpha bungarotoxin alpha conotoxin PeIA bungarotoxin receptor chimeric protein conotoxin nicotinic alpha9alpha10 receptor nicotinic receptor protein subunit receptor subtype recombinant protein snail venom unclassified drug animal cell article binding affinity cholinergic receptor blocking cochlea cochlear nerve concentration response controlled study Conus pergrandis DNA sequence genetic variability hair cell IC 50 matrix assisted laser desorption ionization time of flight mass spectrometry molecular cloning molecular evolution nonhuman oocyte priority journal protein determination protein expression protein function protein localization rat receptor binding receptor sensitivity sequence analysis snail synaptic transmission toxin analysis toxin synthesis wild type Xenopus Acetylcholine Amino Acid Sequence Animals Base Sequence Bungarotoxins Cell Differentiation Cloning, Molecular Conotoxins Dose-Response Relationship, Drug Electrophysiology Ganglia, Spinal Hair Cells, Inner Inhibitory Concentration 50 Kinetics Lymphocytes Molecular Sequence Data Mollusca Neurons Oocytes Peptides Protein Subunits Rats Rats, Sprague-Dawley Receptors, Nicotinic Recombinant Proteins RNA Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Synaptic Transmission Thymus Gland Xenopus Xenopus laevis Conus geographus Conus magus |
| spellingShingle |
Biosynthesis Blood Cells Characterization Cloning Molecular biology Muscle Skin Cochlea Hair cells Thymocytes Xenopus oocytes Biochemistry acetylcholine alpha bungarotoxin alpha conotoxin PeIA bungarotoxin receptor chimeric protein conotoxin nicotinic alpha9alpha10 receptor nicotinic receptor protein subunit receptor subtype recombinant protein snail venom unclassified drug animal cell article binding affinity cholinergic receptor blocking cochlea cochlear nerve concentration response controlled study Conus pergrandis DNA sequence genetic variability hair cell IC 50 matrix assisted laser desorption ionization time of flight mass spectrometry molecular cloning molecular evolution nonhuman oocyte priority journal protein determination protein expression protein function protein localization rat receptor binding receptor sensitivity sequence analysis snail synaptic transmission toxin analysis toxin synthesis wild type Xenopus Acetylcholine Amino Acid Sequence Animals Base Sequence Bungarotoxins Cell Differentiation Cloning, Molecular Conotoxins Dose-Response Relationship, Drug Electrophysiology Ganglia, Spinal Hair Cells, Inner Inhibitory Concentration 50 Kinetics Lymphocytes Molecular Sequence Data Mollusca Neurons Oocytes Peptides Protein Subunits Rats Rats, Sprague-Dawley Receptors, Nicotinic Recombinant Proteins RNA Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Synaptic Transmission Thymus Gland Xenopus Xenopus laevis Conus geographus Conus magus A novel α-conotoxin, PeIA, cloned from Conus pergrandis, discriminates between Rat α9α10 and α7 nicotinic cholinergic receptors |
| topic_facet |
Biosynthesis Blood Cells Characterization Cloning Molecular biology Muscle Skin Cochlea Hair cells Thymocytes Xenopus oocytes Biochemistry acetylcholine alpha bungarotoxin alpha conotoxin PeIA bungarotoxin receptor chimeric protein conotoxin nicotinic alpha9alpha10 receptor nicotinic receptor protein subunit receptor subtype recombinant protein snail venom unclassified drug animal cell article binding affinity cholinergic receptor blocking cochlea cochlear nerve concentration response controlled study Conus pergrandis DNA sequence genetic variability hair cell IC 50 matrix assisted laser desorption ionization time of flight mass spectrometry molecular cloning molecular evolution nonhuman oocyte priority journal protein determination protein expression protein function protein localization rat receptor binding receptor sensitivity sequence analysis snail synaptic transmission toxin analysis toxin synthesis wild type Xenopus Acetylcholine Amino Acid Sequence Animals Base Sequence Bungarotoxins Cell Differentiation Cloning, Molecular Conotoxins Dose-Response Relationship, Drug Electrophysiology Ganglia, Spinal Hair Cells, Inner Inhibitory Concentration 50 Kinetics Lymphocytes Molecular Sequence Data Mollusca Neurons Oocytes Peptides Protein Subunits Rats Rats, Sprague-Dawley Receptors, Nicotinic Recombinant Proteins RNA Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Synaptic Transmission Thymus Gland Xenopus Xenopus laevis Conus geographus Conus magus |
| description |
The α9 and α10 nicotinic cholinergic subunits assemble to form the receptor believed to mediate synaptic transmission between efferent olivocochlear fibers and hair cells of the cochlea, one of the few examples of postsynaptic function for a non-muscle nicotinic acetylcholine receptor (nAChR). However, it has been suggested that the expression profile of α9 and α10 overlaps with that of α7 in the cochlea and in sites such as dorsal root ganglion neurons, peripheral blood lymphocytes, developing thymocytes, and skin. We now report the cloning, total synthesis, and characterization of a novel toxin α-conotoxin PeIA that discriminates between α9α10 and α7 nAChRs. This is the first toxin to be identified from Conus pergrandis, a species found in deep waters of the Western Pacific. α-Conotoxin PeIA displayed a 260-fold higher selectivity for α-bungarotoxin-sensitive α9α10 nAChRs compared with α-bungarotoxin-sensitive α7 receptors. The IC50 of the toxin was 6.9 ± 0.5 nM and 4.4 ± 0.5 nM for recombinant α9α10 and wild-type hair cell nAChRs, respectively. α-Conotoxin PeIA bears high resemblance to α-conotoxins MII and GIC isolated from Conus magus and Conus geographus, respectively. However, neither α-conotoxin MII nor α-conotoxin GIC at concentrations of 10 μM blocked acetylcholine responses elicited in Xenopus oocytes injected with the α9 and α10 subunits. Among neuronal non-α-bungarotoxin- sensitive receptors, α-conotoxin PeIA was also active at α3β2 receptors and chimeric α6/α3β2β3 receptors. α-Conotoxin PeIA represents a novel probe to differentiate responses mediated either through α9α10 or α7 nAChRs in those tissues where both receptors are expressed. © 2005 by The American Society for Biochemistry and Molecular Biology, Inc. |
| title |
A novel α-conotoxin, PeIA, cloned from Conus pergrandis, discriminates between Rat α9α10 and α7 nicotinic cholinergic receptors |
| title_short |
A novel α-conotoxin, PeIA, cloned from Conus pergrandis, discriminates between Rat α9α10 and α7 nicotinic cholinergic receptors |
| title_full |
A novel α-conotoxin, PeIA, cloned from Conus pergrandis, discriminates between Rat α9α10 and α7 nicotinic cholinergic receptors |
| title_fullStr |
A novel α-conotoxin, PeIA, cloned from Conus pergrandis, discriminates between Rat α9α10 and α7 nicotinic cholinergic receptors |
| title_full_unstemmed |
A novel α-conotoxin, PeIA, cloned from Conus pergrandis, discriminates between Rat α9α10 and α7 nicotinic cholinergic receptors |
| title_sort |
novel α-conotoxin, peia, cloned from conus pergrandis, discriminates between rat α9α10 and α7 nicotinic cholinergic receptors |
| publishDate |
2005 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v280_n34_p30107_McIntosh http://hdl.handle.net/20.500.12110/paper_00219258_v280_n34_p30107_McIntosh |
| _version_ |
1768546239785205760 |