Biomedical subjects

Tatyana Polenova

Publications and source records attributed to Tatyana Polenova.

9 recordsLinked to original sources

PubMed2006

Probing local environments in paramagnetic europium-substituted Keggin solids by 31P magic angle spinning NMR spectroscopy.

Paramagnetic Eu-substituted Keggin oxopolytungstates crystallize in different forms, determined by the nature of the counterions. The crystal packing is in turn responsible for the variations in the geometry of paramagnetic Eu sites with respect to the anion core. We probed the paramagnetic environments in a series of Eu-substituted Keggin solids, by 31P magic angle spinning NMR spectroscopy. 31P spinning sideband envelopes are dominated by the electron-nuclear dipolar interaction. For the compounds under investigation, both the magnitude and the asymmetry parameter of the electron-nuclear dipolar coupling tensor are sensitive to the mutual arrangements of paramagnetic Eu sites in the crystal lattice. and also report on the stoichiometry of the anion. The electron-nuclear dipolar coupling tensors were calculated from the crystallographic coordinates and the experimentally determined effective magnetic moments, assuming a point dipole approximation. The computed tensors are in very good agreement with the experimental spectra. Furthermore, the P-Eu distance estimates, accurate to within 0.06-0.12 A, can be obtained directly from the magnitude of the electron-nuclear dipolar coupling. This work demonstrates that 31P MAS NMR spectroscopy is a useful probe for investigating local environments in paramagnetic Keggin solids.

Europium↗

PubMed2006

51V solid-state magic angle spinning NMR spectroscopy of vanadium chloroperoxidase.

We report 51V solid-state NMR spectroscopy of the 67.5-kDa vanadium chloroperoxidase, at 14.1 T. We demonstrate that, despite the low concentration of vanadium sites in the protein (one per molecule, 1 mumol of vanadium spins in the entire sample), the spinning sideband manifold spanning the central and the satellite transitions is readily detectable. The quadrupolar and chemical shift anisotropy tensors have been determined by numerical simulations of the spinning sideband envelopes and the line shapes of the individual spinning sidebands corresponding to the central transition. The observed quadrupolar coupling constant C(Q) of 10.5 +/- 1.5 MHz and chemical shift anisotropy delta(sigma) of -520 +/- 13 ppm are sensitive reporters of the geometric and electronic structure of the vanadium center. Density functional theory calculations of the NMR spectroscopic observables for an extensive series of active site models indicate that the vanadate cofactor is most likely anionic with one axial hydroxo- group and an equatorial plane consisting of one hydroxo- and two oxo- groups. The work reported in this manuscript is the first example of 51V solid-state NMR spectroscopy applied to probe the vanadium center in a protein directly. This approach yields the detailed coordination environment of the metal unavailable from other experimental measurements and is expected to be generally applicable for studies of diamagnetic vanadium sites in metalloproteins.

Binding Sites↗

PubMed2005

Optical spectra of a novel polyoxometalate occluded within modified MCM-41.

The novel polyoxometalate ([(Eu2PW10O38)4(W3O8(H2O)2(OH)4)]22-), also referred to herein as Eu8P4W43, has been immobilized inside the channels of MCM-41 mesoporous molecular sieve material by means of the incipient wetness method. For proper host-guest interaction, amine groups were introduced into the system as a result of an aminosilylation procedure. A stable and integrated Eu8P4W43 polyoxometalate was shown to be formed inside the channels of the modified MCM-41. The products were characterized by XRD, UV-vis absorption, emission, Raman excitation, Raman, and 31P solid-state NMR measurements. Infrared and Raman spectra of the polyoxometalate/MCM-41 composite systems are interpreted as showing spectral shifts due to site induced electrostatic interactions. The photoluminescent behavior of the composite at room temperature indicates a characteristic Eu3+ emission pattern corresponding to 5D0- 7F(J) transitions. A strong photoluminescence suggests the potential utility of the polyoxometalate as a luminescent material.

Aluminum Silicates↗

PubMed2005

Resonance assignments and secondary structure analysis of E. coli thioredoxin by magic angle spinning solid-state NMR spectroscopy.

De novo site-specific 13C and 15N backbone and sidechain resonance assignments are presented for uniformly enriched E. coli thioredoxin, established using two-dimensional homo- and heteronuclear solid-state magic angle spinning NMR correlation spectroscopy. Backbone dihedral angles and secondary structure were derived from the statistical analysis of the secondary chemical shifts, and are in good agreement with solution values for the intact full-length thioredoxin, with the exception of a small number of residues located at the termini of the individual secondary structure elements. A large number of cross-peaks observed in the DARR spectra with long mixing times correspond to the pairs of carbon atoms separated by 4-6 angstroms, suggesting that DARR could be efficiently employed for observation of medium- and long-range correlations. The 108 amino acid residue E. coli thioredoxin is the largest uniformly enriched protein assigned to this degree of completeness by solid-state NMR spectroscopy to date. It is anticipated that with a combination of two-dimensional correlation experiments and high magnetic fields, resonance assignments and secondary structure can be generally derived for other noncrystalline proteins.

Carbon Isotopes↗

PubMed2005

Premicellar complexes of sphingomyelinase mediate enzyme exchange for the stationary phase turnover.

During the steady state reaction progress in the scooting mode with highly processive turnover, Bacillus cereus sphingomyelinase (SMase) remains tightly bound to sphingomyelin (SM) vesicles (Yu et al., Biochim. Biophys. Acta 1583, 121-131, 2002). In this paper, we analyze the kinetics of SMase-catalyzed hydrolysis of SM dispersed in diheptanoylphosphatidyl-choline (DC7PC) micelles. Results show that the resulting decrease in the turnover processivity induces the stationary phase in the reaction progress. The exchange of the bound enzyme (E*) between the vesicle during such reaction progress is mediated via the premicellar complexes (E(i)#) of SMase with DC7PC. Biophysical studies indicate that in E(i)# monodisperse DC7PC is bound to the interface binding surface (i-face) of SMase that is also involved in its binding to micelles or vesicles. In the presence of magnesium, required for the catalytic turnover, three different complexes of SMase with monodisperse DC7PC (E(i)# with i=1, 2, 3) are sequentially formed with Hill coefficients of 3, 4 and 8, respectively. As a result, during the stationary phase reaction progress, the initial rate is linear for an extended period and all the substrate in the reaction mixture is hydrolyzed at the end of the reaction progress. At low mole fraction (X) of total added SM, exchange is rapid and the processive turnover is limited by the steps of the interfacial turnover cycle without becoming microscopically limited by local substrate depletion or enzyme exchange. At high X, less DC7PC will be monodisperse, E(i)# does not form and the turnover becomes limited by slow enzyme exchange. Transferred NOESY enhancement results show that monomeric DC7PC in solution is in a rapid exchange with that bound to E(i)# at a rate comparable to that in micelles. Significance of the exchange and equilibrium properties of the E(i)# complexes for the interpretation of the stationary phase reaction progress is discussed.

Bacillus cereus↗

PubMed2004

Magic angle spinning solid-state NMR spectroscopy for structural studies of protein interfaces. resonance assignments of differentially enriched Escherichia coli thioredoxin reassembled by fragment complementation.

De novo site-specific backbone and side-chain resonance assignments are presented for U-15N(1-73)/U-13C,15N(74-108) reassembly of Escherichia coli thioredoxin by fragment complementation, determined using solid-state magic angle spinning NMR spectroscopy at 17.6 T. Backbone dihedral angles and secondary structure predicted from the statistical analysis of 13C and 15N chemical shifts are in general agreement with solution values for the intact full-length thioredoxin, confirming that the secondary structure is retained in the reassembled complex prepared as a poly(ethylene glycol) precipitate. The differential labeling of complementary thioredoxin fragments introduced in this work is expected to be beneficial for high-resolution structural studies of protein interfaces formed by protein assemblies by solid-state NMR spectroscopy.

Amino Acid Sequence↗

PubMed2004

51V magic angle spinning NMR spectroscopy of Keggin anions [PVnW12-nO40](3+n)-: effect of countercation and vanadium substitution on fine structure constants.

Vanadium environments in Keggin oxopolytungstates were characterized by (51)V solid-state MAS NMR spectroscopy. (C(4)H(9))(4)N(+)-, K(+)-, Cs(+)-, as well as mixed Na(+)/Cs(+)- salts of the mono-, di-, and trivanadium substituted oxotungstates, [VW(11)O(40)](4-), [V(2)W(10)O(40)](5-), and [V(3)W(9)O(40)](6-), have been prepared as microcrystalline and crystalline solids. Solid-state NMR spectra report on the local environment of the vanadium site in these Keggin ions via their anisotropic quadrupolar and chemical-shielding interactions. These (51)V fine structure constants in the solid state are determined by the number of vanadium atoms present in the oxoanion core. Surprisingly, the quadrupolar anisotropy tensors do not depend to any significant extent on the nature of the countercations. On the other hand, the chemical-shielding anisotropy tensors, as well as the isotropic chemical shifts, display large variations as a function of the cationic environment. This information can be used as a probe of the local cationic environment in the vanadium-substituted Keggin solids.

Anions↗

PubMed2003

51V magic angle spinning NMR spectroscopy of six-coordinate Lindqvist oxoanions: a sensitive probe for the electronic environment in vanadium-containing polyoxometalates. Counterions dictate the 51V fine structure constants in polyoxometalate solids.

Geometric and electronic environments of vanadium have been addressed by (51)V magic angle spinning NMR spectroscopy of six-coordinated polyoxometalate solids. (C(4)H(9))(4)N(+) and mixed Na(+)/Cs(+) salts of the Lindqvist-type mono- and divanadium-substituted oxotungstates, [VW(5)O(19)](3-) and [V(2)W(4)O(19)](4-), have been prepared as microcrystalline and crystalline solids. The solid-state NMR spectra reflect the details of the local environment of the vanadium site in these hexametalate solids via the anisotropic quadrupolar and chemical shielding interactions. Remarkably, these (51)V fine structure constants in the solid state are dictated by the nature and geometry of the countercations. Electrostatic calculations of the electric field gradients at the vanadium atoms have been performed. Experimental trends are well reproduced with the simple electrostatic model, and explain the sensitivity of the anisotropic NMR parameters to the changes in the cationic environment at the vanadium site.

Journal Article↗

PubMed2003

51V solid-state magic angle spinning NMR spectroscopy and DFT studies of oxovanadium(V) complexes mimicking the active site of vanadium haloperoxidases.

A series of 11 oxovanadium(V) complexes mimicking the active site of vanadium haloperoxidases have been investigated by (51)V magic angle spinning NMR spectroscopy and density functional theory (DFT). The MAS spectra are dominated by the anisotropic quadrupolar and chemical shielding interactions; for these compounds, C(Q) ranges from 3 to 8 MHz, and delta(sigma) is in the range 340-730 ppm. The quadrupolar coupling and chemical shielding tensors as well as their relative orientations have been determined by numerical simulations of the spectra. The spectroscopic NMR observables appear to be very sensitive to the details of the electronic and geometric environment of the vanadium center in these complexes. For the four crystallographically characterized compounds from the series, the quadrupolar and chemical shielding anisotropies were computed at the DFT level using two different basis sets, and the calculated tensors were in general agreement with the experimental solid-state NMR data. A combination of (51)V solid-state NMR and computational methods is thus beneficial for investigation of the electrostatic and geometric environment in diamagnetic vanadium systems with moderate quadrupolar anisotropies.

Algorithms↗