• Title/Summary/Keyword: $^{31}P$ NMR spectroscopy

Search Result 46, Processing Time 0.023 seconds

Observation of Asymmetry amongst Nucleotide Binding Sites of F1-ATPase of Escherichia coli by 31P NMR Spectroscopy

  • Jun, Nam-Kung;Sohn, Joon-Hyung;Yeh, Byung-Il;Choi, Jong-Whan;Kim, Hyun-Won
    • Bulletin of the Korean Chemical Society
    • /
    • v.32 no.2
    • /
    • pp.531-535
    • /
    • 2011
  • It was regarded that the $^{31}P$ resonances of inherent nucleotides in $F_1$-ATPase (EF1), as large as 380KDa, could not be observed by $^{31}P$ NMR spectroscopy. However, our $^{31}P$ NMR spectroscopy could differentiate between different nucleotide binding sites on EF1 from Escherichia coli. When EF1 was prepared in the absence of $Mg^{2+}$, EF1 contained only ADP. Multiple $^{31}P$ resonances from $\beta$-phosphates of ADP bound to the EF1 were observed from the enzyme prepared without $Mg^{2+}$, suggesting asymmetry or flexibility amongst nucleotide binding sites. $^{31}P$ resonances from enzyme bound ATP could be observed only from EF1, when the enzyme was prepared in the presence of $Mg^{2+}$. This $Mg^{2+}$ dependent ATP binding was very tight that, once bound, nucleotide could not be removed even after removal of $Mg^{2+}$. $^{31}P$ NMR proved to be a valuable tool for investigating phosphorous related enzymes.

31P NMR Spectroscopy Revealed Adenylate kinase-like Activity and Phosphotransferase-like Activity from F1-ATPase of Escherichia coli

  • Kim, Hyun-Won
    • Bulletin of the Korean Chemical Society
    • /
    • v.32 no.1
    • /
    • pp.183-185
    • /
    • 2011
  • Adenylate kinase-like activity and phosphotransferase-like activity from $F_1$-ATPase of Escherichia coli was revealed by $^{31}P$ NMR spectroscopy. Incubation of F1-ATPase with ADP in the presence of $Mg^{2+}$ shows the appearance of $^{31}P$ resonances from AMP and Pi, suggesting generation of AMP and ATP by adenylate kinase-like activity and the subsequent hydrolysis to Pi. Incubation of $F_1$-ATPase with ADP in the presence of methanol shows additional peak from methyl phosphate, suggesting phosphotransferase-like activity of $F_1$-ATPase. Both adenylate kinase-like activity and phosphotransferase-like activity has not been reported from $F_1$-ATPase of Escherichia coli. $^{31}P$ NMR could be a valuable tool for the investigation of phosphorous related enzyme.

Localized In Vivo $^{31}P$ NMR Studies on Rabbit Skeletal Muscle Tissue from Premortem to Postmortem Period

  • Choe, Bo-Young;Kim, Sung-Eun;Lee, Hyoung-Koo;Suh, Tae-Suk;Lee, Heung-Kyu;Shinn, Kyung-Sub
    • Journal of the Korean Magnetic Resonance Society
    • /
    • v.3 no.1
    • /
    • pp.1-11
    • /
    • 1999
  • Localized in vivo 31P NMR spectroscopy was applied to evaluate the postmortem catabolism of high energy phosphates in rabbit skeletal muscle tissue. In the premortem processes all of the important high energy phosphate metabolites were characterized, and particularly phosphocreatine (PCr) resonance signal was the strongest. In the immediate phases of the postmortem processes the signal intensities of PCr, phosphomonoesters (PME), phosphodiesters(PDE), $\alpha$-, $\beta$- and ${\gamma}$-adenosine triphosphate (ATP) resonance began to decrease while the signal intensity of inorganic phosphorus (Pi) resonance began to increase. The present study suggests that localized in vivo 31P NMR spectroscopy may provide more precise biochemical information of the early postmortem period based on the metabolic alterations of phosphate. The unique ability of localized in vivo 31P NMR spectroscopy to offer noninvasive information about tissue biochemistry in animals as well as human may have an impact on thanatochronology and medicolegal science.

  • PDF

The Status of Guanine Nucleotides in Taxol-Stabilized Microtubules Probed by 31P CPMAS NMR Spectroscopy

  • Ferdous, Taslima;Lee, Sang-Hak;Yeo, Kwon-Joo;Paik, Youn-Kee
    • Journal of the Korean Magnetic Resonance Society
    • /
    • v.15 no.2
    • /
    • pp.104-114
    • /
    • 2011
  • Rapid exchange and hydrolysis of the tubulin-bound guanine nucleotides have been known to govern the dynamics of microtubules. However, the instability and low concentration have made it difficult for the microtubule-bound GTP to be observed directly. In this study, we circumvent these problems by lyophilization and using cross-polarization techniques. $^{31}P$ NMR signals were detected from the tubulin-bound GTP in microtubules for the first time. Analysis of the $^{31}P$ CPMAS NMR spectrum indicates that GTP hydrolysis was delayed by the presence of taxol.

Characterization of C-P Lyase gene cluster by in vivo $^{31}$ P-NMR spectroscopy

  • Lee, Ki-Sung;Kwak, In-Young
    • Journal of Microbiology
    • /
    • v.33 no.4
    • /
    • pp.328-333
    • /
    • 1995
  • $\^$31/ P-NMR experiment was performed to detect phophonates (Pn) utilization and degradation in the several different C-P lyase mutants of E. coli and in E. aerogenes and the recombinants. The relative peak intensity (RPI) for the standard samples of 0.5 mM methylphosphonate (MPn) and 1.0 mM aminoethylphosphonate in glucose-MOPS medium showed 0.5 : 1.0 ratio. In the case of BW14329 (.DELTA.phnC-P, .delta.phoA), RPI did not change significantly after 24 hrs culturing, which means it nearly could not utilize Pn. In vivo $\^$31/ P-NMR spectrum of E. aerogens (BWKL 16627) during 3 hrs starvation showed two intense peaks at 0-2 ppm and at near-10 ppm which indicate intracellular orthophosphate (Pi) and pyrophosphate (PPi), respectively. Both of them might be released by degradation of inorganic polyphosphate pool. When MPn is supplied to the medium as an unique P source, Pi content in the cell has the constant, but PPi seems to be slightly decreased. Recombinants (BWKL 16954) grew slower than E. aerogenes in the glucose-MOPS media with various P sources. In vivo $\^$31/ P-NMR spectrum of recombinant did not show any intense signal in the cell. Surprisingly, under the cultivation adding with MPn, a few intense peaks in the region of Pi AND phospate monoester were detected.

  • PDF

A Study of Phosphate Adsorption on Kaolinite by $^{31}$P NMP Spectroscopy ($^{31}$P NMR을 이용한 카올리나이트에 흡착된 인산염의 연구)

  • 김영규
    • Journal of the Mineralogical Society of Korea
    • /
    • v.13 no.4
    • /
    • pp.186-195
    • /
    • 2000
  • To study phosphate adsorption on kaolinite, $^{31}$ P MAS NMR(magic angle spinning nuclear magnetic resonance spectroscopy)has been used for kaolinite reacted in 0.1 M phosphate solutions at pH’s from 3 to 11. There are at least 3 different forms of phosphate on kaolinite. One is the phosphate physically adsorbed on kaolinite surface (outer-sphere complexes) or species left after vacuum-filtering. The second is the phosphate adsorbed by ligand exchange (inner-sphere complexes), and the third is Al-phosphate precipitates which are pH dependent. Most of the inner-spherer complexes and surface precipitates are mainly on hydroxided Al(aluminol) rather than hydroxided Si(silanol). These are pertinent with the results obtained from the phosphate adsorption experiments on silica gel and ${\gamma}$-Al$_2$O$_3$ as model compounds, respectively. The two peaks with more negative chemical shifts(more shielded) than the ortho-phosphate peak (positive chemical shift) are assigned to be the inner-sphere complexes and surface precipitates. The $^{31}$ P chemical shifts of the Al-phosphate precipitates are more negative than those of inner-sphere complexes at a given pH due to the larger number of P-O-Al linkages per tetrahedron. The chemical shifts of both the inner-sphere complexes and surface precipitates are more negative than those of inner-sphere complexes at a given pH due to the larger number of P-O-Al linkages per tetrahedron. The chemical shifts of both the inner-sphere complexes and surface precipitates become progressively less shielded with increasing pH. For the inner-sphere complexes, decreasing phosphate protonation combined with peak averaging by rapid proton exchange among phosphate tetrahedra with different numbers of protons is though to be the reason for the peak change. The decreasing shielding with increasing pH for surface precipitates is probably due to the decreasing average number of P-O-Al linkages per tetrahedron combined with decreasing protonation like inner-sphere complexes.

  • PDF

Characterization of the Catalytic Heteropoly Compounds using Solid-state NMR

  • Kim, Y;Lee, W
    • Journal of the Korean Magnetic Resonance Society
    • /
    • v.1 no.1
    • /
    • pp.45-58
    • /
    • 1997
  • Heteropoly compounds, H3PMo12O40, CsxH3-xPMo12O40, and vanadium containing heteropoly compound were characterized by Solid-state broad line 1H MAS NMR, 31P MAS NMR, and High Speed MAS 51V NMR spectroscopy of quadrupolar nuclei. The effects of calcination, dehydration, and the number of protons on the structure of heteropoly compounds were studied. The results of this study demonstrate that these Solid-state NMR techniques are very useful tools to study heteropoly compounds.

  • PDF

Hard Tissue Analysis of NMR after Fluoride Administration

  • Kim, Hye-Young;Nam, Seoul-Hee;Han, Man-Seok
    • Journal of Magnetics
    • /
    • v.21 no.4
    • /
    • pp.599-602
    • /
    • 2016
  • Fluoride (F) is an important element for the mineralization of body tissues. The purpose of this study was to administer fluoride prenatally to rats to evaluate its beneficial concentration for rat bone using microstructural analysis, to analyze its effect on the bone structure, and to evaluate the effect of its transfer through rat placenta. Fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance spectrometry (NMR) were performed. The $^{19}F$ NMR and $^{31}P$ NMR signals suggested the existence of fluoride ions in the apatite lattice because the signals were caused by the fluoride ions that were coupled to the phosphate atoms and were affected in the phosphate phases other than the element phases in the apatite. Consequently, if it was not affected too much, the desirable concentration of prenatal fluoride treatment could have a helpful effect on the bone crystal structure through placental fluoride transfer.