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Guanosine 5 triphosphate

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Guanosine-5′-triphosphate is a nucleoside triphosphate. It is the activated form of the nucleoside guanosine and serves as a substrate for various enzymes involved in cellular processes.

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Lab products found in correlation

Dithiothreitol (dtt), by Merck Group (3 mentions) Adenosine 5 triphosphate, by Merck Group (3 mentions) Triton x 100, by Merck Group (2 mentions) Propidium iodide, by Merck Group (2 mentions) Ethylene glycol tetraacetic acid (egta), by Merck Group (2 mentions) Taxol, by Merck Group (2 mentions) Uridine 5 triphosphate, by Merck Group (2 mentions) Bovine serum albumin (bsa), by Merck Group (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Pfu ultra dna polymerase, by Agilent Technologies (1 mentions) Depc treated water, by Quality Biological (1 mentions) T7 dna ligase, by New England Biolabs (1 mentions) 2 deoxyguanosine 5 triphosphate dgtp, by Promega (1 mentions) 2 deoxyuridine 5 triphosphate dutp, by Roche (1 mentions) T7 rna polymerase, by New England Biolabs (1 mentions) T4 polynucleotide kinase, by New England Biolabs (1 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions) Phenylmethanesulfonyl fluoride, by Merck Group (1 mentions) Phosphatase inhibitor, by Thermo Fisher Scientific (1 mentions) Sodium chloride (nacl), by Merck Group (1 mentions)

Spelling variants of this product

Guanosine (93 citations) Guanosine triphosphate (13 citations) Guanosine 5′-triphosphate sodium salt hydrate (6 citations) Sodium guanosine-13C10 5′-triphosphate (4 citations) Guanosine 5′-[γ-thio]triphosphate tetralithium salt (cold GTPγS) (2 citations) Guanosine-5′-triphosphate (GTP) (2 citations)

12 protocols using guanosine 5 triphosphate

1

Radiolabeled Nucleotide Synthesis Protocol

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2′-Deoxyguanosine-5′-triphosphate (dGTP) was obtained from Promega, 2′-deoxyuridine-5′-triphosphate (dUTP) was obtained from Roche, tritiated 2′-deoxyuridine-5′-triphosphate tetraammonium salt ([5–3H] dUTP) and tritiated 2′-deoxyguanosine-5′-triphosphate tetraammonium salt ([8-3H] dGTP) were from Moravek Biochemicals, 2′-deoxyguanosine-5′-[α-thio] triphosphate lithium salt (dGTPαS) was from ChemCyte, guanosine-5′-triphosphate was from Sigma-Aldrich, and C18-reversed phase thin layer chromatography (TLC) plates were purchased from Macherey-Nagel. DEPC-treated water was obtained from Quality Biological, Inc. T7 RNA polymerase, T4 polynucleotide kinase, T7 DNA ligase and restriction enzymes were obtained from New England Biolabs. Pfu Ultra DNA polymerase was obtained from Agilent Technologies.
Seamon K.J., Sun Z., Shlyakhtenko L.S., Lyubchenko Y.L, & Stivers J.T. (2015). SAMHD1 is a single-stranded nucleic acid binding protein with no active site-associated nuclease activity. Nucleic Acids Research, 43(13), 6486-6499.
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2

Comprehensive Biochemical Assay Protocol

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Gold (III) chloride trihydrate (HAuCl4.3H2O), dichlorofluorescein diacetate (DCFH-DA), Rhodamine 123, protease inhibitor cocktail, phenylmethanesulfonyl fluoride (PMSF), propidium iodide (PI), formaldehyde, Taxol, ethidium bromide, guanosine-5′-triphosphate (GTP), glutamate, piperazine- N, N′-bis (2-ethane sulfonic acid) (Pipes), magnesium sulfate (MgSO4), ethylene glycol tetraacetic acid (EGTA), vinblastine, dithiothreitol (DTT), iodoacetamide (IAA), ammonium bicarbonate (ABC), trifluoroacetic acid (TFA), formic acid, sodium chloride, Triton X-100, and sodium deoxycholate were procured from Sigma (St. Louis, MO). Dulbecco’s Modified Eagle’s Medium (DMEM), fetal bovine serum (FBS), trypsin-EDTA (0.25%) solution, penicillin, streptomycin, bovine serum albumin (BSA), horse serum, (3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT), ribonuclease A, dimethyl sulfoxide (DMSO), acetonitrile, and HPLC-grade water were purchased from Himedia, India. Tryptone, Tris buffer, acridine orange, and crystal violet were from Sisco Research Laboratories (SRL, Bangalore, India). Phosphatase inhibitor, trypsin-protease, and Prolong Gold anti-fade reagent were obtained from Thermo Scientific (Waltham, Massachusetts). Urea was obtained from Rankem, India. All other chemicals and solvents were also of analytical grade and highest purity.
Nirmala J.G, & Lopus M. (2019). Tryptone-stabilized gold nanoparticles induce unipolar clustering of supernumerary centrosomes and G1 arrest in triple-negative breast cancer cells. Scientific Reports, 9, 19126.
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3

GTP Kinetics Analysis Protocol

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Guanosine 5′-triphosphate (GTP), S-adenosyl-L-methionine (SAM), dithiothreitol (DTT), and sodium dithionite were purchased from Sigma-Aldrich. [3′-D]GTP was prepared as described previously.11 (link) Nonlinear least-squares fitting of kinetic data was carried out using KaleidaGraph software (Synergy Software, Reading, PA). Anaerobic experiments were carried out in an UNIlab workstation glovebox (MBaun, Stratham, NH) maintained at 10 ± 2 °C with an O2 concentration <0.1 ppm. All HPLC experiments were performed on a Hitachi L-2130 pump equipped with an L-2455 diode array detector, an L-2485 fluorescence detector, an L-2200 autosampler, and an ODS Hypersil C18 column (Thermo Scientific) housed in an L-2300 column oven maintained at 40 °C. UV–vis absorption spectra were determined using the U-3900 UV–vis ratio recording double-beam spectrometer (Hitachi). Synthetic peptides were purchased from Genscript.
Hover B.M, & Yokoyama K. (2015). C-Terminal Glycine-Gated Radical Initiation by GTP 3′,8-Cyclase in the Molybdenum Cofactor Biosynthesis. Journal of the American Chemical Society, 137(9), 3352-3359.
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4

Purification and Analysis of GTPases

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Guanosine-5′-triphosphate (GTP), guanosine-5′-diphosphate (GDP), guanosine-5′-(β,γ-imino) triphosphate (GDPNP) and Sephacryl S-300 HR resin were purchased from Sigma. HisPur Ni-NTA Superflow Agarose resin was purchased from Thermo Scientific. TALON resin was purchased from Clontech. Malachite Green dye was purchased from Fisher Scientific.
Carlson M.A., Haddad B.G., Weis A.J., Blackwood C.S., Shelton C.D., Wuerth M.E., Walter J.D, & Spiegel PC J.r. (2017). Ribosomal Protein L7/L12 is Required for GTPase Translation Factors EF-G, RF3 and IF2 to Bind in their GTP State to 70S Ribosomes. The FEBS journal, 284(11), 1631-1643.
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5

Nucleotide Exchange of K-RAS and Mutants

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K-RAS and its oncogenic mutants at a concentration of 100 μM were incubated in buffer D supplemented with 10 mM ethylenediaminetetraacetic acid (EDTA) and either 2 mM GTP (guanosine 5′-triphosphate, Sigma-Aldrich) or dGTP (2′-deoxyguanosine 5′-triphosphate, Sigma-Aldrich) for 2 h at 4 °C. The nucleotide exchange reaction mixture was then supplemented with 10 mM magnesium chloride and incubated for an additional 1 h at 4 °C. Excess nucleotide was removed using a centrifugal buffer exchange device (Micro Bio-Spin 6 Column, Bio-Rad). To fully exchange GDP with GTP, this procedure was repeated once more. The nucleotide-exchanged sample was then supplemented with either GTP or dGTP at a final concentration of 2 mM, added to a Slide-A-Lyzer MINI device (2000 Da MWCO, Thermo), and dialyzed against 200 mM ammonium acetate overnight at 4 °C.
Moghadamchargari Z., Huddleston J., Shirzadeh M., Zheng X., Clemmer D.E., Raushel F.M., Russell D.H, & Laganowsky A. (2019). Intrinsic GTPase Activity of K-RAS Monitored by Native Mass Spectrometry. Biochemistry, 58(31), 3396-3405.
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6

Cytoskeleton Disruption Assay with Small Molecules

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Paclitaxel, vinblastine sulphate, podophyllotoxin, colchicine, 5,5′‐dithiobis‐2‐nitrobenzoic acid (DTNB), sulforhodamine B (SRB), Hoechst 33342, guanosine 5′‐triphosphate (GTP), propidium iodide, EGTA, MgCl2, piperazine‐N,N′‐bis (2‐ethanesulphonic acid) (PIPES), mouse monoclonal anti‐α‐tubulin IgG and FITC‐conjugated anti‐mouse IgG, fluorescein isothiocyanate isomer (FITC) and dimethylformamide were purchased from Sigma‐Aldrich (St. Louis, MO, USA). Foetal bovine serum (FBS) and Alexa Fluor 568‐conjugated anti‐mouse IgG were purchased from Invitrogen (Thermo Scientific, Massachusetts, USA). Acridine orange (AO), hydroxysuccinimide, minimal essential medium (MEM), cell culture tested antibiotic solution, and phosphate‐buffered saline (PBS) were purchased from HiMedia (Mumbai, India). Dichloromethane, n‐hexane, hydroxylamine hydrochloride and triethylamine were purchased from Merck, India. 1‐Ethyl‐3‐(3‐dimethylaminopropyl) carbodiimide hydrochloride was purchased from SRL chemicals. All other reagents used in the study were of analytical grade.
Ashraf S.M., Sebastian J, & Rathinasamy K. (2018). Zerumbone, a cyclic sesquiterpene, exerts antimitotic activity in HeLa cells through tubulin binding and exhibits synergistic activity with vinblastine and paclitaxel. Cell Proliferation, 52(2), e12558.
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7

Neonatal Rat Cardiomyocyte Isolation

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Adenosine 5′-triphosphate (ATP), guanosine 5′-triphosphate (GTP), uridine 5′-triphosphate (UTP), cytidine 5′-triphosphate (CTP), and oligomycin were purchased from Sigma-Aldrich (St Louis, MO, USA). Imaging compatible Nunc™ Lab-Tek™ II 8-well chamber slides (Cat#: 155409PK), Hoechst 33,342 (Cat#: H3570), Tubulin Tracker™ Green (Oregon Green™ 488 Taxol, Cat#: T34075), neonatal rat cardiomyocyte isolation enzymes, supplements, and primary cell isolation medias were purchased form Thermo Fisher Scientific (Waltham, MA). Neonatal rat hearts were generously donated from FIU’s animal facility. All the other chemicals were of analytical reagent grade and were used as received without further purification.
Liu W., Zhu X., Mozneb M., Nagahara L., Hu T.Y, & Li C.Z. (2021). Lighting up ATP in cells and tissues using a simple aptamer-based fluorescent probe. Mikrochimica Acta, 188(10), 352.
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8

Quantification of Nucleotide Levels

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All reagents were of analytical grade. Bovine serum albumin (BSA), malachite green, adenosine 5′ -triphosphate (ATP), adenosine 5′ -diphosphate (ADP), cytidine 5′-triphosphate disodium salt (CTP), adenosine 5′ -monophosphate (AMP), uridine 5’-triphosphate (UTP), uridine 5’-diphosphate (UDP), guanosine-5’-triphosphate (GTP), phosphate-buffered saline (DPBS), 4-(2-hydroxyethyl)-1-piperazineetahnesulfonic acid (HEPES), ammonium molybdate, Triton X-100, phenylmethylsulphonyl fluoride (PMSF), pyruvate kinase, phosphoenol-pyruvate (PEP), luciferase, coenzyme A and P1,P5-Di (adenosine-5´) pentaphosphate pentasodium salt (Ap5A) were purchased from Sigma-Aldrich (St Louis, MO, USA). D-luciferin was purchased from Molecular Probes Inc. (Eugene, OR, USA).
Saffioti N.A., Alvarez C.L., Bazzi Z., Gentilini M.V., Gondolesi G.E., Schwarzbaum P.J, & Schachter J. (2023). Dynamic recycling of extracellular ATP in human epithelial intestinal cells. PLOS Computational Biology, 19(6), e1011196.
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9

Microtubule Enrichment and Analysis

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This was performed as described previously28 (link) with minor modifications. Briefly, cells synchronized and enriched in mitotis were lysed at 4°C in 100 mM 1,4-piperazinediethanesulfonic acid (PIPES), at pH 6.8, 1 mM MgCl2, 2 mM ethylene glycol tetra acetic acid (EGTA) and 1% Triton X-100, and spun at 13 000 × g for 30 min. To the supernatant thus obtained, purified tubulin (Cytoskeleton, Denver, CO, USA; 4 μg), dithiothreitol (1 mM), guanosine-5′-triphosphate (GTP; 1 mM) and taxol (Sigma-Aldrich) (10 μM) were added to cleared lysates, and incubated for 1 h at 37°C. Nocodazole (100 μM) was added as a negative control. Lysates were layered over a 20% sucrose cushion in the above buffer and spun at 48 000 r.p.m. for 1 h at room temperature (rotor TLA 120.1, Beckman Coulter). Microtubule pellets were collected after removing lysate and cushion, bound proteins were separated by SDS-PAGE and analyzed by western blotting.
Srivastava D, & Chakrabarti O. (2014). Mahogunin-mediated α-tubulin ubiquitination via noncanonical K6 linkage regulates microtubule stability and mitotic spindle orientation. Cell Death & Disease, 5(2), e1064-.
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10

Electrochemical Biosensing of Neurochemicals

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Dopamine (DA), adenosine (AD), adenosine triphosphate (ATP), guanosine (GN), and guanosine-5′-triphosphate (GTP) were purchased from Sigma-Aldrich (St. Louis, MO, USA). All neurochemical stock solutions in this study were 10 mM, dissolved in 0.1 M HCl, and stored at 4 °C. Tris buffer at a pH of 7.4 was used to dilute stock solutions to produce daily working solutions (1 μM DA, 5 μM AD, ATP, GN, and GTP) for testing electrodes. Tris buffer includes 15 mM Tris, 1.25 mM NaH2PO4, 2.0 mM Na2SO4, 3.25 mM KCl, 140 mM NaCl, 1.2 mM CaCl2 dihydrate, and 1.2 mM MgCl2. All solutions were made from deionized water (Milli-Q, Millipore, Billerica MA).
Syeed A.J., Li Y., Ostertag B.J., Brown J.W, & Ross A.E. (2022). Nanostructured carbon-fiber surfaces for improved neurochemical detection. Faraday discussions, 233, 336-353.
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