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GRIN2A protein, human
GRIN2A protein, human
The GRIN2A protein is a subunit of the N-methyl-D-aspartate (NMDA) receptor, which is a type of ionotropic glutamate receptor that plays a key role in synaptic transmission and neuroplasticity in the central nervous system.
The GRIN2A gene encodes this protein, which is essential for proper brain development and function.
Mutations in GRIN2A have been linked to various neurological disorders, including epilepsy, intellectual disability, and autism spectrum disorders.
Researchers can explore the GRIN2A protein using PubCompare.ai's AI-powered platform to discover optimized research protocols, improve reproducibility, and identify the best products for their GRIN2A protein research.
This streamlined workflow can help scientists get the most out of their experiements and advance our understanding of this important protein.
The GRIN2A gene encodes this protein, which is essential for proper brain development and function.
Mutations in GRIN2A have been linked to various neurological disorders, including epilepsy, intellectual disability, and autism spectrum disorders.
Researchers can explore the GRIN2A protein using PubCompare.ai's AI-powered platform to discover optimized research protocols, improve reproducibility, and identify the best products for their GRIN2A protein research.
This streamlined workflow can help scientists get the most out of their experiements and advance our understanding of this important protein.
Most cited protocols related to «GRIN2A protein, human»
Animal Care Committees
Animals
Animals, Laboratory
Antibodies
Brain
Cannula
GRIN2A protein, human
Immunohistochemistry
Lentivirinae
Males
Memory
Mice, House
Mice, Knockout
Microscopy, Fluorescence
paraform
Pentobarbital
Rabbits
Seahorses
Sucrose
Technique, Dilution
X-ray diffraction data sets were collected at the Advanced Light Source on beamlines 8.2.1 and 5.0.2. Diffraction sets were indexed, integrated, and scaled by XDS57 or HKL200058 together with the microdiffraction assembly method59 (link). The best diffraction data for Data set 1 were derived from merging data from three crystals60 (link). A single crystal was used for Data set 2. Structure 1 was determined by molecular replacement with Phaser61 (link) using the isolated Xenopus/rat GluN1/GluN2B ATD domains in complex with Ro25-6981 (PDB code 3QEM)24 (link) and the isolated rat/rat GluN1/GluN2A LBD (PDB code 2A5T)32 (link) structures as search probes. The molecular replacement solutions were robust with the highest best log likelihood gain and translation function Z-score of 3071.7 and 31.9, respectively. Initial maps were improved by density modification62 (link). A partial model of the transmembrane domain was manually built into ‘omit’ style electron density maps. Cycles of manual model building and crystallographic refinement were carried out using the computer graphic program Coot63 and the crystallographic refinement software package Phenix64 (link). During the course of model building and refinement, the amino acid sequence and corresponding structure within the ATDs and LBDs were adjusted to the Xenopus amino acid sequences. The model was refined to a nominal resolution of 3.7 Å with reasonable R-factors. Structure 2 derived from Data set 2 was solved by molecular replacement using Structure 1 as a search probe. Upon inspection of electron density maps, density for the pore loops was visible, along with additional residues in the other TM segments. The final Structure 2 was obtained by cycles of manual model building and crystallographic refinement, as described above. Stereochemistry of the model was evaluated by MolProbity 65 (link), pore dimensions were estimated using HOLE66 (link) and figures were created using Pymol67 . Important information on the qualities of the structures is provided in Supplementary Information .
Amino Acid Sequence
Ataxia Telangiectasia
Crystallography
Electrons
GRIN2A protein, human
GRIN2B protein, human
Lafora Disease
Light
Microtubule-Associated Proteins
R Factors
X-Ray Diffraction
Xenopus laevis
Amino Acids
Amino Acid Sequence
Arginine
DNA, Complementary
GRIN2A protein, human
GRIN2B protein, human
Homo sapiens
N-Methyl-D-Aspartate Receptors
Oocytes
Open Reading Frames
Protein Subunits
Untranslated Regions
Xenopus laevis
Amino Acids
Amino Acid Sequence
bacteriophage T7 RNA polymerase
Cells
Chimera
Cloning Vectors
Codon, Terminator
DNA, Complementary
GRIN2A protein, human
GRIN2B protein, human
Helix (Snails)
Oocytes
Peptides
Promega
Proteins
Protein Subunits
Signal Peptides
Staphylococcal Protein A
Xenopus laevis
Antibodies
GRIN2A protein, human
Synapsin I
Synaptophysin
Tubulin
Most recents protocols related to «GRIN2A protein, human»
Triheteromeric receptor
constructs were generated using rat GluN1 and GluN2A with modified
C-terminal peptide tags as previously described.72 (link) Briefly, C-terminal peptide tags were generated from leucine
zipper motifs found in GABAB1 (referred to as C1) and GABAB2 (referred to as C2). These tags were placed downstream of
a synthetic helical linker and upstream of a KKTN endoplasmic reticulum
retention signal.113 (link)−115 (link) The tag was introduced in frame and in place
of the stop codon at the GluN2A C-terminal tail to make 2AC1 and 2AC2. A chimeric GluN2B subunit was constructed in
which the 2B carboxyl tail after residue 838 was replaced by the GluN2A
carboxyl tail and C-terminal-linker-C1 or -C2-ER retention motifs
to make 2BAC1 and 2BAC2.72 (link) The C1 and C2 leucine zipper motifs can form a coiled-coil
structure that masks the KKTN retention motif and allows for expression
of only triheteromeric receptors on the cell surface. Recordings were
taken at pH 7.4.
Measurement of “escape” currents
was used to assess the efficiency of the peptide tags which control
surface expression. Our average escape currents were typically less
than 10% and this was considered an acceptable threshold. Currents
were estimated using pairs of mutations (GluN2A-R518K,T690I and GluN2B-R519K,T691I)
that render the agonist binding domain incapable of binding glutamate,
and therefore unable to pass current.
constructs were generated using rat GluN1 and GluN2A with modified
C-terminal peptide tags as previously described.72 (link) Briefly, C-terminal peptide tags were generated from leucine
zipper motifs found in GABAB1 (referred to as C1) and GABAB2 (referred to as C2). These tags were placed downstream of
a synthetic helical linker and upstream of a KKTN endoplasmic reticulum
retention signal.113 (link)−115 (link) The tag was introduced in frame and in place
of the stop codon at the GluN2A C-terminal tail to make 2AC1 and 2AC2. A chimeric GluN2B subunit was constructed in
which the 2B carboxyl tail after residue 838 was replaced by the GluN2A
carboxyl tail and C-terminal-linker-C1 or -C2-ER retention motifs
to make 2BAC1 and 2BAC2.72 (link) The C1 and C2 leucine zipper motifs can form a coiled-coil
structure that masks the KKTN retention motif and allows for expression
of only triheteromeric receptors on the cell surface. Recordings were
taken at pH 7.4.
Measurement of “escape” currents
was used to assess the efficiency of the peptide tags which control
surface expression. Our average escape currents were typically less
than 10% and this was considered an acceptable threshold. Currents
were estimated using pairs of mutations (GluN2A-R518K,T690I and GluN2B-R519K,T691I)
that render the agonist binding domain incapable of binding glutamate,
and therefore unable to pass current.
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Cells
Chimera
Codon, Terminator
Glutamate
GRIN2A protein, human
GRIN2B protein, human
Helix (Snails)
Leucine Zippers
Mutation
Peptides
Protein Subunits
Reading Frames
Retention (Psychology)
Tail
tyrosyl-alanyl-glycine
Rat cDNA encoding GluN1-1a,
(GluN1, RefSeq NP_058706), GluN2A (NP_036705), GluN2B (NP_036706),
GluN2C (NP_036707), and GluN2D (NP_073634) were obtained from Drs.
S. Heinemann (Salk Institute), S. Nakanishi (Kyoto University), and
P. Seeburg (University of Heidelberg). cRNA was transcribed in vitro
from linearized plasmids containing NMDAR cDNAs according to the manufacturer’s
instructions (mMessage mMachine, Ambion; Thermo Fisher Scientific,
Waltham, MA). NMDARs were expressed in X. laevis oocytes following microinjection of 3–5 ng of the GluN1 subunit
cRNA and 7–10 ng of GluN2B subunit cRNA in 50 nL of RNAse free
water as previously described.112 (link) Oocytes
were incubated in Barth’s solution at 18 °C, and recordings
were made 2–7 days after the injections at room temperature
using two two-electrode voltage clamp amplifiers at a holding potential
of −40 mV.
Oocytes were perfused with a solution of 90
mM NaCl, 1 mM KCl, 10 mM HEPES, and 0.5 mM BaCl2, and the
pH was adjusted to 7.4 using 1 M NaOH. 10 μM of EDTA was added
to chelate contaminant divalent ions such as Zn2+. Oocytes
were placed in a dual-track plexiglass recording chamber that was
assumed to be at a reference potential of 0 mV. The glass microelectrodes
were filled with 300 mM KCl for the voltage electrode, and 3 M KCl
for the current electrode. Bath clamps communicating across silver
chloride wires were placed into each side of the recording chamber.
The IC50 data was obtained by applying 100 μM glutamate
and 30 μM glycine, followed by the application of glutamate
and glycine plus increasing concentrations of the test compound up
to 30 μM. Current responses of less than 50 nA were not included.
The level of inhibition was calculated as a percent of the initial
glutamate response, averaged across all oocytes from a single frog.
Each experiment used 6–7 oocytes from the same frog. The results
from these experiments were pooled and fitted to the equation, where minimum
is the residual percent response
at saturating concentration of the test compound and nH is a slope factor for the steepness of the inhibition curve.
(GluN1, RefSeq NP_058706), GluN2A (NP_036705), GluN2B (NP_036706),
GluN2C (NP_036707), and GluN2D (NP_073634) were obtained from Drs.
S. Heinemann (Salk Institute), S. Nakanishi (Kyoto University), and
P. Seeburg (University of Heidelberg). cRNA was transcribed in vitro
from linearized plasmids containing NMDAR cDNAs according to the manufacturer’s
instructions (mMessage mMachine, Ambion; Thermo Fisher Scientific,
Waltham, MA). NMDARs were expressed in X. laevis oocytes following microinjection of 3–5 ng of the GluN1 subunit
cRNA and 7–10 ng of GluN2B subunit cRNA in 50 nL of RNAse free
water as previously described.112 (link) Oocytes
were incubated in Barth’s solution at 18 °C, and recordings
were made 2–7 days after the injections at room temperature
using two two-electrode voltage clamp amplifiers at a holding potential
of −40 mV.
Oocytes were perfused with a solution of 90
mM NaCl, 1 mM KCl, 10 mM HEPES, and 0.5 mM BaCl2, and the
pH was adjusted to 7.4 using 1 M NaOH. 10 μM of EDTA was added
to chelate contaminant divalent ions such as Zn2+. Oocytes
were placed in a dual-track plexiglass recording chamber that was
assumed to be at a reference potential of 0 mV. The glass microelectrodes
were filled with 300 mM KCl for the voltage electrode, and 3 M KCl
for the current electrode. Bath clamps communicating across silver
chloride wires were placed into each side of the recording chamber.
The IC50 data was obtained by applying 100 μM glutamate
and 30 μM glycine, followed by the application of glutamate
and glycine plus increasing concentrations of the test compound up
to 30 μM. Current responses of less than 50 nA were not included.
The level of inhibition was calculated as a percent of the initial
glutamate response, averaged across all oocytes from a single frog.
Each experiment used 6–7 oocytes from the same frog. The results
from these experiments were pooled and fitted to the equation, where minimum
is the residual percent response
at saturating concentration of the test compound and nH is a slope factor for the steepness of the inhibition curve.
Full text: Click here
A-factor (Streptomyces)
barium chloride
Bath
Complementary RNA
DNA, Complementary
Edetic Acid
Endoribonucleases
factor A
Glycine
GRIN2A protein, human
GRIN2B protein, human
HEPES
Ions
Microinjections
N-Methyl-D-Aspartate Receptors
Oocytes
Plasmids
Plexiglas
Protein Subunits
Psychological Inhibition
Rana
Sodium Chloride
Xenopus laevis
The crystal structures of MAPK3, SRC, MAPK1, NMDAR2B and NMDAR2A were obtained from the RCSB Protein Data Bank (https://www.rcsb.org/ , accessed on 18 August 2022, PDB codes: 4QTB, 2H8H, 4QTA, 7EU8 and 7EU7). All original ligands (including (3R)-1-(2-oxo-2-{4-[4-(pyrimidin-2-yl)phenyl]piperazin-1-yl}ethyl)-N-[3-(pyridin-4-yl)-2H-indazol-5-yl]pyrrolidine-3-carboxamide, N-(5-CHLORO-1,3-BENZODIOXOL-4-YL)-7-[2-(4-METHYLPIPERAZIN-1-YL)ETHOXY]-5-(TETRAHYDRO-2H-PYRAN-4-YLOXY)QUINAZOLIN-4-AMINE, (3R)-1-(2-oxo-2-{4-[4-(pyrimidin-2-yl)phenyl]piperazin-1-yl}ethyl)-N-[3-(pyridin-4-yl)-2H-indazol-5-yl]pyrrolidine-3-carboxamide, and S-ketamine) and water molecules were removed, and hydrogen atoms and charges were added to the macromolecules. The three-dimensional structures of Gelsemium alkaloids downloaded from the PubChem database and optimized by Chem 3D Pro15.0 were used as the ligand. Molecular docking was finalized by AutoDock Vina [27 (link)]. The size of the gridbox was fixed to 40 × 40 × 40 angstroms, with a spacing of 0.375 angstrom. All the parameters of the genetic algorithm (GA) were set to the default values. The conformers with the lowest binding energy were selected for analysis.
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Alkaloids
Gelsemium
GRIN2A protein, human
GRIN2B protein, human
Hydrogen
Indazoles
Ketamine
Ligands
MAPK1 protein, human
mitogen-activated protein kinase 3, human
N-methylacetamide-oxotremorine M
Piperazine
Pyrans
pyrrolidine
quinazolin-4-amine
Acetylcholine receptor a-subunit R-97-116, dopamine D1 receptor domain–3, 4 and 5, dopamine D2 receptor DRD2 E1.1 and E1.2, N-methyl-D-aspartate receptors NR2A and NR2B, amyloid-b peptide 1-42, and enteric nerve ribonuclear polypeptide A were synthesized by Bio-Synthesis (Lewisville, TX, USA), and tau protein recombinant brain-derived neurotropic factor (BDNF) was purchased from R&D Systems (Minneapolis, MN, USA).
Full text: Click here
Amyloid Proteins
Anabolism
Brain
Cholinergic Receptors
Dopamine D1 Receptor
Dopamine D2 Receptor
DRD2 protein, human
GRIN2A protein, human
GRIN2B protein, human
MAPT protein, human
N-Methyl-D-Aspartate Receptors
Nervousness
Peptides
Polypeptides
Protein Subunits
Homogenates were linearized in 1× laemmli buffer (Bio-Rad Laboratories, Hercules, CA, USA) with 5% β-mercaptoethanol (Abcam, Cambridge, UK) at 99 °C for 10 min. Homogenate containing 50 μg protein was loaded in each lane. Proteins were separated by 10% SDS polyacrylamide gel (Bio-Rad Laboratories, Hercules, CA, USA) and transferred to polyvinylidene fluoride (PVDF) membranes (Bio-Rad Laboratories, Hercules, CA, USA). Non-phosphorylated protein targets were blocked by 5% non-fat dry milk powder (Bio-Rad Laboratories, Hercules, CA, USA) and phospho-protein targets were blocked by 5% bovine serum albumin (Sigma-Aldrich, St. Loius, MO, USA) in the Tris-HCl buffer (pH 8.0). After 1 h blocking, membranes were incubated overnight with primary antibodies in blocking solution with 0.05% Tween−20. The following primary antibodies were used: rabbit anti-PSD95 (1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-GluA1 (1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-GluN2A (1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-GluN2B (1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-phospho-GluN2A1246 (1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-phospho-GluN2B1472 (1:1000, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-adiponectin R1 (1:1000, Abcam, Cambridge, UK), goat anti-adiponectin R2 (1:1000, Novus Biologicals, Centennial, CO, USA), rabbit anti-α-tubulin (1:1000, Cell Signaling Technology, Danvers, MA, USA), mouse anti-β-actin (1:5000, Invitrogen, Waltham, MA, USA). After washing, membranes were incubated with the corresponding secondary HRP-linked antibodies for 1 h: goat anti-rabbit IgG (1:1000, Cell Signaling Technology, Danvers, MA, USA), goat anti-mouse IgG (1:1000, Cell Signaling Technology, Danvers, MA, USA), and donkey anti-goat IgG (1:1000, Invitrogen, Waltham, MA, USA), and developed by Clarity Western ECL Substrate (Bio-Rad Laboratories, Hercules, CA, USA) for chemiluminescence detection and documented by a transilluminator (Bio-Rad Laboratories, Hercules, CA, USA). Band intensities were subjected to densitometric analysis in the Image Lab software (Bio-Rad Laboratories, Hercules, CA, USA).
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2-Mercaptoethanol
Actins
Adiponectin
alpha-Tubulin
anti-IgG
Antibodies
Biological Factors
Cardiac Arrest
Chemiluminescence
Densitometry
Equus asinus
Goat
GRIN2A protein, human
GRIN2B protein, human
Laemmli buffer
Milk, Cow's
Mus
Novus
polyacrylamide gels
polyvinylidene fluoride
Powder
Proteins
Protein Targeting, Cellular
Rabbits
Serum Albumin, Bovine
Tissue, Membrane
Tromethamine
Tween 20
Top products related to «GRIN2A protein, human»
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GluN2A is a subunit of the NMDA receptor, a type of ionotropic glutamate receptor found in the central nervous system. It plays a key role in the regulation of synaptic function and plasticity. The GluN2A subunit contributes to the receptor's properties, including ion permeability and channel kinetics.
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β-actin is a protein that is found in all eukaryotic cells and is involved in the structure and function of the cytoskeleton. It is a key component of the actin filaments that make up the cytoskeleton and plays a critical role in cell motility, cell division, and other cellular processes.
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GluN2B is a subunit of the N-methyl-D-aspartate (NMDA) receptor, which is an ionotropic glutamate receptor that plays a key role in synaptic transmission and neuronal plasticity. The GluN2B subunit is essential for the proper functioning and regulation of the NMDA receptor.
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PVDF membranes are a type of laboratory equipment used for a variety of applications. They are made from polyvinylidene fluoride (PVDF), a durable and chemically resistant material. PVDF membranes are known for their high mechanical strength, thermal stability, and resistance to a wide range of chemicals. They are commonly used in various filtration, separation, and analysis processes in scientific and research settings.
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TRIzol reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components designed for the isolation of total RNA, DNA, and proteins from a variety of biological samples. The reagent maintains the integrity of the RNA while disrupting cells and dissolving cell components.
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Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for efficient and reliable delivery of nucleic acids, such as plasmid DNA and small interfering RNA (siRNA), into a wide range of eukaryotic cell types. It facilitates the formation of complexes between the nucleic acid and the lipid components, which can then be introduced into cells to enable gene expression or gene silencing studies.
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Quantity One software is a powerful and versatile tool for analyzing and quantifying data from gel electrophoresis and imaging experiments. It provides a suite of analytical tools for researchers to accurately measure and compare the size, intensity, and other properties of bands or spots in their samples.
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Anti-GluN2A is a lab equipment product used to detect and quantify the GluN2A subunit of the N-methyl-D-aspartate (NMDA) receptor. It is a specific antibody that can be used in various research applications, such as Western blotting, immunohistochemistry, and immunoprecipitation, to study the expression and localization of the GluN2A subunit.
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GluA1 is a subunit of the AMPA-type glutamate receptor, which is a ligand-gated ion channel involved in fast excitatory neurotransmission in the central nervous system. This product can be used for research purposes to study the structure and function of AMPA receptors.
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More about "GRIN2A protein, human"
The GRIN2A gene encodes the GluN2A subunit of the N-methyl-D-aspartate (NMDA) receptor, a key player in synaptic transmission and neuroplasticity in the central nervous system.
This ionotropic glutamate receptor is essential for proper brain development and function.
Mutations in GRIN2A have been linked to various neurological disorders, including epilepsy, intellectual disability, and autism spectrum disorders.
Researchers can explore the GRIN2A protein using PubCompare.ai's AI-powered platform to discover optimized research protocols, improve reproducibility, and identify the best products for their experiments.
This includes comparing literature, preprints, and patents to find the most effective approaches.
For GRIN2A protein studies, common experimental techniques involve Western blotting with Anti-GluN2A antibodies, immunoprecipitation, and quantification using Quantity One software.
Samples may be prepared using TRIzol reagent and Lipofectamine 2000 transfection.
Downstream analysis can include assessments of β-actin as a loading control and measurements of GluA1 expression.
PubCompare.ai's AI-driven comparisons can help scientists streamline their GRIN2A protein research workflow, improve reproducibility, and advance our understanding of this important neurological target.
Leveraging this platform can maximize the impact of experiments and accelerate discoveries related to GRIN2A and associated neurological disorders.
This ionotropic glutamate receptor is essential for proper brain development and function.
Mutations in GRIN2A have been linked to various neurological disorders, including epilepsy, intellectual disability, and autism spectrum disorders.
Researchers can explore the GRIN2A protein using PubCompare.ai's AI-powered platform to discover optimized research protocols, improve reproducibility, and identify the best products for their experiments.
This includes comparing literature, preprints, and patents to find the most effective approaches.
For GRIN2A protein studies, common experimental techniques involve Western blotting with Anti-GluN2A antibodies, immunoprecipitation, and quantification using Quantity One software.
Samples may be prepared using TRIzol reagent and Lipofectamine 2000 transfection.
Downstream analysis can include assessments of β-actin as a loading control and measurements of GluA1 expression.
PubCompare.ai's AI-driven comparisons can help scientists streamline their GRIN2A protein research workflow, improve reproducibility, and advance our understanding of this important neurological target.
Leveraging this platform can maximize the impact of experiments and accelerate discoveries related to GRIN2A and associated neurological disorders.