scholarly journals A novel method for the synthesis of nucleoside triphosphates labelled with inorganic [32P]phosphate specifically in the β-position

1977 ◽  
Vol 161 (3) ◽  
pp. 615-617
Author(s):  
K A Abraham
Keyword(s):  
Phosphate Group ◽  
Polynucleotide Kinase ◽  
Nucleoside Triphosphates ◽  
Novel Method ◽  
Coupled Reaction ◽  
Phosphate Groups

A method was developed for the introduction of [32p]Pi specifically into the beta-position of ATP and GTP. The method is based on two separate reactions involving (a) phosphorolysis of poly(A) or poly(G) [Soreq, Nudel, Salomon, Revel & Littauer (1974) J. Mol Biol. 88, 233-245] in the presence of [32P]Pi and (b) conversion of the labelled diphosphate into the corresponding triphosphate by transferring the active phosphate group from 1,3-diphosphoglycerate in a coupled reaction as decribed by Glynn & Chappell [(1964) Biochem. J. 90, 147-149]. Radioactivity in the beta- and gamma-phosphate groups of the labelled triphosphate was measured by using polynucleotide kinase. No detectable radioactivity was found in the gamma-phosphate group. The suitability of this method for the synthesis of other nucleoside triphosphates specifically labelled in the beta-position is discussed.

The first 3′:5′-cyclic nucleotide–amino acid complex:L-His–cIMP

2012 ◽  
Vol 68 (8) ◽  
pp. o311-o316 ◽  
Author(s):  
Katarzyna Ślepokura
Keyword(s):  
Crystal Structure ◽  
Amino Acid ◽  
Cyclic Nucleotide ◽  
Phosphate Group ◽  
Specific Amino Acid ◽  
Π Stacking ◽  
Cyclic Phosphate ◽  
Nucleotide Recognition ◽  
Group Play ◽  
Phosphate Groups

In the crystal structure of the L-His–cIMP complex,i.e.L-histidinium inosine 3′:5′-cyclic phosphate [systematic name: 5-(2-amino-2-carboxyethyl)-1H-imidazol-3-ium 7-hydroxy-2-oxo-6-(6-oxo-6,9-dihydro-1H-purin-9-yl)-4a,6,7,7a-tetrahydro-4H-1,3,5,2λ5-furo[3,2-d][1,3,2λ5]dioxaphosphinin-2-olate], C6H10N3O2+·C10H10N4O7P−, the Hoogsteen edge of the hypoxanthine (Hyp) base of cIMP and the Hyp face are engaged in specific amino acid–nucleotide (His...cIMP) recognition,i.e.by abutting edge-to-edge and by π–π stacking, respectively. The Watson–Crick edge of Hyp and the cIMP phosphate group play a role in nonspecific His...cIMP contacts. The interactions between the cIMP anions (anti/C3′–endo/trans–gauche/chair conformers) are realized mainly between riboses and phosphate groups. The results for this L-His–cIMP complex, compared with those for the previously reported solvated L-His–IMP crystal structure, indicate a different nature of amino acid–nucleotide recognition and interactions upon the 3′:5′-cyclization of the nucleotide phosphate group.

The Position of Phosphate Groups in the Phosphonomannan of Hansenula capsulata, as Determined by Carbon-13 Magnetic Resonance Spectroscopy

1973 ◽  
Vol 51 (13) ◽  
pp. 2105-2109 ◽  
Author(s):  
Philip A. J. Gorin
Keyword(s):  
Magnetic Resonance ◽  
Magnetic Resonance Spectroscopy ◽  
Spectral Data ◽  
Resonance Spectrum ◽  
Phosphate Group ◽  
Resonance Spectroscopy ◽  
Previous Assignment ◽  
Phosphate Groups

The carbon-13 magnetic resonance spectrum of a phosphate of 2-O-β-D-mannopyranosyl-α,β-D-mannose was compared with that of the unphosphorylated disaccharide. The positions and number of the signals of C-5's which are coupled to phosphorus-31, together with other spectral data show that the phosphate group is on C-6 of the reducing end-unit (2, Fig. 1). This differs from a previous assignment (1), thus necessitating a revision (4) to the structure 3 proposed for the parent Hansenula capsulata phosphonomannan.

A Novel Method for the Preparation of Nucleoside Triphosphates from Activated Nucleoside Phosphoramidates

2004 ◽  
Vol 6 (13) ◽  
pp. 2257-2260 ◽  
Author(s):  
Weidong Wu ◽  
Caren L. Freel Meyers ◽  
Richard F. Borch
Keyword(s):  
Nucleoside Triphosphates ◽  
Novel Method ◽  
Nucleoside Phosphoramidates

Adsorption between TC-stabilized AuNPs and the phosphate group: application of the PTP1B activity assay

The Analyst ◽  
2015 ◽  
Vol 140 (23) ◽  
pp. 8017-8022 ◽  
Author(s):  
Jun Lv ◽  
Xiaonan Wang ◽  
Yuanyuan Zhang ◽  
Defeng Li ◽  
Juan Zhang ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
Magnetic Nanoparticles ◽  
Colorimetric Method ◽  
Phosphate Group ◽  
Activity Assay ◽  
Phosphate Groups

Based on the adsorption between tetracycline (TC) and phosphate groups, a general colorimetric method is explored in this work by using TC-stabilized gold nanoparticles (TC/AuNPs) and 4-aminophenyl phosphate-functionalized Fe3O4 magnetic nanoparticles (APP/MNPs).

scholarly journals Aqueous Contact Ion Pairs of Phosphate Groups with Na+, Ca2+ and Mg2+ – Structural Discrimination by Femtosecond Infrared Spectroscopy and Molecular Dynamics Simulations

2020 ◽  
Vol 234 (7-9) ◽  
pp. 1453-1474 ◽  
Author(s):  
Benjamin P. Fingerhut ◽  
Jakob Schauss ◽  
Achintya Kundu ◽  
Thomas Elsaesser
Keyword(s):  
Ion Pairs ◽  
Model System ◽  
Ion Pair ◽  
Pair Formation ◽  
Phosphate Group ◽  
Phosphate Ion ◽  
2D Ir ◽  
Dna And Rna ◽  
Contact Ion Pairs ◽  
Phosphate Groups

AbstractThe extent of contact and solvent shared ion pairs of phosphate groups with Na+, Ca2+ and Mg2+ ions in aqueous environment and their relevance for the stability of polyanionic DNA and RNA structures is highly debated. Employing the asymmetric phosphate stretching vibration of dimethyl phosphate (DMP), a model system of the sugar-phosphate backbone of DNA and RNA, we present linear infrared, femtosecond infrared pump-probe and absorptive 2D-IR spectra that report on contact ion pair formation via the presence of blue shifted spectral signatures. Compared to the linear infrared spectra, the nonlinear spectra reveal contact ion pairs with increased sensitivity because the spectra accentuate differences in peak frequency, transition dipole moment strength, and excited state lifetime. The experimental results are corroborated by long time scale MD simulations, benchmarked by density functional simulations on phosphate-ion-water clusters. The microscopic interpretation reveals subtle structural differences of ion pairs formed by the phosphate group and the ions Na+, Ca2+ and Mg2+. Intricate properties of the solvation shell around the phosphate group and the ion are essential to explain the experimental observations. The present work addresses a challenging to probe topic with the help of a model system and establishes new experimental data of contact ion pair formation, thereby underlining the potential of nonlinear 2D-IR spectroscopy as an analytical probe of phosphate-ion interactions in complex biological systems.

scholarly journals Phosphorylation of arylsulphatase A occurs through multiple interactions with the UDP-N-acetylglucosamine-1-phosphotransferase proximal and distal to its retrieval site by the KDEL receptor

1999 ◽  
Vol 340 (3) ◽  
pp. 729-736 ◽  
Author(s):  
Frank DITTMER ◽  
Kurt VON FIGURA
Keyword(s):  
Lysosomal Enzymes ◽  
Inhibitory Effect ◽  
Phosphate Group ◽  
Site Specific ◽  
Arylsulphatase A ◽  
Kdel Receptor ◽  
Multiple Interactions ◽  
Phosphate Groups ◽  
Retention Signal ◽  
Ionophore Monensin

Phosphorylation of oligosaccharides of the lysosomal enzyme arylsulphatase A (ASA), which accumulate in the secretions of cells that mis-sort most of the newly synthesized lysosomal enzymes due to a deficiency of mannose 6-phosphate receptors, was found to be site specific. ASA residing within the secretory route of these cells contains about one third of the incorporated [2-3H]mannose in phosphorylated oligosaccharides. Oligosaccharides carrying two phosphate groups are almost 2-fold less frequent than those with one phosphate group and only a few of the phosphate groups are uncovered. Addition of a KDEL (Lys-Asp-Glu-Leu) retention signal prolongs the residence time of ASA within the secretory route 6-fold, but does not result in more efficient phosphorylation. In contrast, more than 90% of the [2-3H]mannose incorporated into secreted ASA (with or without a KDEL retention signal) is present in phosphorylated oligosaccharides. Those with two phosphate groups are almost twice as frequent as those with one phosphate group and most of the phosphate groups are uncovered. Thus, ASA receives N-acetylglucosamine 1-phosphate groups in a sequential manner at two or more sites located within the secretory route proximal and distal to the site where ASA is retrieved by the KDEL receptor, i.e. proximal to the trans-Golgi. At each of these sites up to two N-acetylglucosamine 1-phosphate groups can be added to a single oligosaccharide. Of several drugs known to inhibit transit of ASA through the secretory route only the ionophore monensin had a major inhibitory effect on phosphorylation, uncovering and sialylation.

Nanostructured and Electroactive Hybrid Films Containing Microcrystalline Cellulose Modified with the Phosphate Group: Synthesis and Characterization

2016 ◽  
Vol 869 ◽  
pp. 840-845
Author(s):  
Paulo Ronaldo Sousa Teixeira ◽  
Ana Siqueira do Nascimento Marreiro ◽  
José Regilmar Teixeira da Silva ◽  
Emanuel Airton de Oliveira Farias ◽  
Natália de Araujo Dionisio ◽  
...  
Keyword(s):  
Microcrystalline Cellulose ◽  
Self Assembly ◽  
Phosphate Group ◽  
Synthesis And Characterization ◽  
Layer By Layer ◽  
Preparation Process ◽  
Hybrid Films ◽  
Assembly Technique ◽  
Phosphate Groups ◽  
Modified Cellulose

Nanostructured and electroactive hybrid films containing Microcrystalline Cellulose (MC) modified with the phosphate group (MCPO4) and polyaniline (PANI) were prepared using the Layer-by-Layer (LbL) self-assembly technique. In the preparation process of the films cellulose was dispersed in the solution of PANI and the film with PVS/PANI(MCPO4) structure was immobilized on ITO substrate. In order to investigate the influence of the phosphate group on the electrochemical behavior of the film, films were also prepared replacing the modified cellulose (MCPO4) by unmodified cellulose (MC), forming the PVS/PANI(MC) films. Subsequently, all films were characterized by cyclic voltammetry (CV), and the results showed the redox processes characteristic of PANI, but the presence of CMPO4 promotes an increase in the values of current density observed for the PVS/PANI(MCPO4) film when compared to the PVS/PANI(MC) film. This is probably due to a self-doping process of the polymer in the presence of phosphate groups.

scholarly journals Detecting the Site of Phosphorylation in Phosphopeptides without Loss of Phosphate Group Using MALDI TOF Mass Spectrometry

2008 ◽  
Vol 3 ◽  
pp. ACI.S497 ◽  
Author(s):  
Medicharla V. Jagannadham ◽  
Ramakrishnan Nagaraj
Keyword(s):  
Mass Spectrometry ◽  
Negative Ions ◽  
Maldi Mass Spectrometry ◽  
Phosphate Group ◽  
Maldi Tof Mass Spectrometry ◽  
Linear Mode ◽  
Maldi Tof ◽  
Phosphorylated Peptides ◽  
Delayed Extraction ◽  
Phosphate Groups

Phosphopeptides with one and four phosphate groups were characterized by MALDI mass spectrometry. The molecular ion of monophosphopeptide could be detected both as positive and negative ions by MALDI TOF with delayed extraction (DE) and in the reflector mode. The tetraphospho peptide could be detected in linear mode. When MS/MS spectra of the monophospho peptides were obtained in a MALDI TOF TOF instrument by CID, b and y ions with the intact phosphate group were observed, in addition the b and y ions without the phosphate group. Our study indicates that it is possible to detect phosphorylated peptides with out the loss of phosphate group by MALDI TOF as well as MALDI TOF TOF instruments with delayed extraction and in the reflector mode.

scholarly journals Phosphatidylinositol 3,4,5-trisphosphate is formed from phosphatidylinositol 4,5-bisphosphate in thrombin-stimulated platelets

1994 ◽  
Vol 301 (2) ◽  
pp. 415-420 ◽  
Author(s):  
A N Carter ◽  
R Huang ◽  
A Sorisky ◽  
C P Downes ◽  
S E Rittenhouse
Keyword(s):  
Specific Activity ◽  
Specific Radioactivity ◽  
Phosphate Group ◽  
Thrombin Receptor ◽  
Dependent Manner ◽  
Metabolic Route ◽  
Metabolic Sequence ◽  
Phosphoinositide 3 Kinase ◽  
The Individual ◽  
Phosphate Groups

Platelets accumulate PtdIns(3,4,5)P3 and PtdIns(3,4)P2 in response to thrombin and thrombin-receptor-directed peptide in a GTP-dependent manner. These phosphoinositides are considered to be mediators of signaling events in a variety of cells. We have examined the metabolic route by which PtdIns(3,4,5)P3 and PtdIns(3,4)P2 are synthesized by briefly (10 min) incubating platelets with high activities of [32P]Pi, followed by 20 or 60 s exposure to thrombin, and analysing the relative radioactivities of the individual phosphate groups in the resulting labelled PtdIns(3,4,5)P3 and PtdIns(3,4)P2. The phosphate group possessing the highest specific activity under such non-equilibrium labelling conditions indicates the last one added in a metabolic sequence. The thrombin-stimulated rate of labelling of PtdIns(3,4)P2 was significantly slower than that of PtdIns(3,4,5)P3. Increased labelled PtdIns3P was not detected within 60 s. The measured relative radioactivities decreased in the order 3 > 5 > 4 >> 1 for PtdIns(3,4,5)P3 and 3 > 4 >> 1 for PtdIns(3,4)P2. On the basis of the results of both rate-of-labelling and specific radioactivity analyses we conclude that PtdIns(3,4,5)Pa is formed by 3-OH phosphorylation of PtdIns(4,5)P2, whereas PtdIns(3,4)P2, may be formed by 3-OH phosphorylation of PtdIns4P and/or dephosphorylation of PtdIns(3,4,5)P3. These findings point to the activation of phosphoinositide 3-kinase as a critical receptor-regulated step in thrombin-stimulated platelets.

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