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61.
A new type of spin diffusion, cross-relaxation driven spin diffusion (CRDSD), is investigated using (15)N NMR on a N-acetyl-L-valyl-L-leucine (NAVL) single crystal under stationary condition. A two-dimensional (2D) pulse sequence that correlates the chemical shifts of (15)N nuclei, with a radio-frequency spin lock on the (15)N channel during the mixing time, is used to observe CRDSD. Experimental results obtained using CRDSD, rf-driven spin diffusion, and proton driven spin diffusion approaches on the NAVL single crystal are compared. Our experimental results suggest that the (15)N spin diffusion rate can be enhanced by about 1000 times using CRDSD than by the normal proton driven spin diffusion. Interestingly, the required spin-locking rf field strength for CRDSD is much lower than that used for the rf-driven spin diffusion experiments. The cross-peak patterns observed in 2D (15)N-(15)N correlation spectra using CRDSD and RFDSD are very different as they arise from different spin-spin interactions. A detailed theory describing CRDSD and RFDSD processes is also presented using a thermodynamic model. The speedy spin diffusion process rendered by the CRDSD approach will be useful to assign resonances from a uniformly (15)N or (13)C labeled proteins and peptides, particularly in aligned samples. 相似文献
62.
A Minimal Functional Complex of Cytochrome P450 and FBD of Cytochrome P450 Reductase in Nanodiscs
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Elke Prade Mukesh Mahajan Sang‐Choul Im Meng Zhang Katherine A. Gentry Prof. G. M. Anantharamaiah Prof. Lucy Waskell Prof. Ayyalusamy Ramamoorthy 《Angewandte Chemie (International ed. in English)》2018,57(28):8458-8462
Structural interactions that enable electron transfer to cytochrome‐P450 (CYP450) from its redox partner CYP450‐reductase (CPR) are a vital prerequisite for its catalytic mechanism. The first structural model for the membrane‐bound functional complex to reveal interactions between the full‐length CYP450 and a minimal domain of CPR is now reported. The results suggest that anchorage of the proteins in a lipid bilayer is a minimal requirement for CYP450 catalytic function. Akin to cytochrome‐b5 (cyt‐b5), Arg 125 on the C‐helix of CYP450s is found to be important for effective electron transfer, thus supporting the competitive behavior of redox partners for CYP450s. A general approach is presented to study protein–protein interactions combining the use of nanodiscs with NMR spectroscopy and SAXS. Linking structural details to the mechanism will help unravel the xenobiotic metabolism of diverse microsomal CYP450s in their native environment and facilitate the design of new drug entities. 相似文献
63.
Thirupathi Ravula Nathaniel Z. Hardin Sudheer Kumar Ramadugu Sarah J. Cox Prof. Ayyalusamy Ramamoorthy 《Angewandte Chemie (International ed. in English)》2018,57(5):1342-1345
Polymer lipid nanodiscs are an invaluable system for structural and functional studies of membrane proteins in their near‐native environment. Despite the recent advances in the development and usage of polymer lipid nanodisc systems, lack of control over size and poor tolerance to pH and divalent metal ions are major limitations for further applications. A facile modification of a low‐molecular‐weight styrene maleic acid copolymer is demonstrated to form monodispersed lipid bilayer nanodiscs that show ultra‐stability towards divalent metal ion concentration over a pH range of 2.5 to 10. The macro‐nanodiscs (>20 nm diameter) show magnetic alignment properties that can be exploited for high‐resolution structural studies of membrane proteins and amyloid proteins using solid‐state NMR techniques. The new polymer, SMA‐QA, nanodisc is a robust membrane mimetic tool that offers significant advantages over currently reported nanodisc systems. 相似文献
64.
Knowledge of the orientation of the nitrogen-15 chemical shift anisotropy (CSA) tensor is critical for a variety of experiments that provide information on protein structure and dynamics in the solid and solution states. Unfortunately, the methods available for determining the orientation of the CSA tensor experimentally have inherent limitations. Rotation studies of a single crystal provide complete information but are tedious and limited in applicability. Solid-state NMR studies on powder samples can be applied to a greater range of samples but suffer from ambiguities in the results obtained. Density functional gauge-including-atomic-orbitals (GIAO) calculations of the orientations of (15)N CSA tensors in peptides are presented here as an independent source of confirmation for these studies. A comparison of the calculated (15)N CSA orientations with the available experimental values from single-crystal and powder studies shows excellent agreement after a partial, constrained optimization of some of the crystal structures used in the calculation. The results from this study suggest that the orientation as well as the magnitudes of (15)N CSA tensors may vary from molecule to molecule. The calculated alpha(N) angle varies from 0 degrees to 24 degrees with the majority in the 10 degrees to 20 degrees range and the beta(N) angle varies from 17 degrees to 24 degrees in good agreement with most of the solid-state NMR experimental results. Hydrogen bonding is shown to have negligible effect on the orientation of (15)N CSA tensor in accordance with recent theoretical predictions. Furthermore, it is demonstrated that the orientation of the (15)N CSA can be calculated accurately with much smaller basis sets than is needed to calculate the chemical shift, suggesting that the routine application of ab initio calculations to the determination of (15)N CSA tensor orientations in large biomolecules might be possible. 相似文献
65.
66.
Shilpa Kammaradi Sanjeeva Swathi Korrapati Chandrasekhar B. Nair P. V. Subba Rao Phani Kumar Pullela U. Vijayalakshmi Ramamoorthy Siva 《Journal of fluorescence》2014,24(4):1297-1306
Donor –linker –acceptor (DSSA) is a concept in fluorescence chemistry with acceptor being a fluorescent compound (FRET) or quencher. The DSSA probes used to measure thiol levels in vitro and in vivo. The reduction potential of these dyes are in the range of ?0.60 V, much lower than the best thiol reductant reported in literature, the DTT (?0.33 V). DSSA disulphide having an unusually low reduction potential compared to the typical thiol reductants is a puzzle. Secondly, DSSA probes have a cyclized rhodamine ring as acceptor which does not have any spectral overlap with fluorescein, but quenches its absorbance and fluorescence. To understand the structural features of DSSA probes, we have synthesized DSSANa and DSSAOr. The calculated reduction potential of these dyes suggest that DSSA probes have an alternate mechanism from the FRET based quenching, namely hydrophobic interaction or dye to dye quenching. The standard reduction potential change with increasing complexity and steric hindrance of the molecule is small, suggesting that ultra- low Eo’ has no contribution from the disulphide linker and is based on structural interactions between fluorescein and cyclized rhodamine. Our results help to understand the DSSA probe quenching mechanism and provide ways to design fluorescent probes. 相似文献
67.
Selectivity in photocatalysis by particulate semiconductors 总被引:1,自引:0,他引:1
Chockalingam Karunakaran Ramamoorthy Dhanalakshmi 《Central European Journal of Chemistry》2009,7(1):134-137
TiO2, Fe2O3, CuO, ZnO, ZnS, Nb2O5, MoO3, CdO, CdS, Sb2O3, CeO2, HgO, Pb2O3, PbO2 and Bi2O3 microparticles exhibit band gap excitation with UV-A light but they are selective to photodegrade phenols. While TiO2 anatase and ZnO photocatalyze the degradation of phenol, o-aminophenol, m-aminophenol, p-aminophenol, o-chlorophenol, m-chlorophenol,
p-chlorophenol, o-nitrophenol, p-nitrophenol, o-cresol, m-cresol, p-cresol, catechol, resorcinol and quinol, MoO3 does not photodegrade any of the fifteen phenols. Fe2O3, CuO, ZnS, Nb2O5, CdO, CdS, Sb2O3, CeO2, HgO, Pb2O3, PbO2 and Bi2O3 are selective in photodegrading the fifteen phenols; however, the phenols get adsorbed over all sixteen particulate semiconductors.
相似文献
68.
Nguyen KT Soong R Lm SC Waskell L Ramamoorthy A Chen Z 《Journal of the American Chemical Society》2010,132(43):15112-15115
In addition to providing a semipermeable barrier that protects a cell from harmful stimuli, lipid membranes occupy a central role in hosting a variety of biological processes, including cellular communications and membrane protein functions. Most importantly, protein-membrane interactions are implicated in a variety of diseases and therefore many analytical techniques were developed to study the basis of these interactions and their influence on the molecular architecture of the cell membrane. In this study, sum frequency generation (SFG) vibrational spectroscopy is used to investigate the spontaneous membrane insertion process of cytochrome b(5) and its mutants. Experimental results show a significant difference in the membrane insertion and orientation properties of these proteins, which can be correlated with their functional differences. In particular, our results correlate the nonfunctional property of a mutant cytochrome b(5) with its inability to insert into the lipid bilayer. The approach reported in this study could be used as a potential rapid screening tool in measuring the topology of membrane proteins as well as interactions of biomolecules with lipid bilayers in situ. 相似文献
69.
Tris-(8-hydroxyquinoline)indium (Inq3) thin films, one of the metal chelates used in organic electroluminescent (EL) devices, were deposited by thermal evaporation process on glass substrates and were exposed to light for various time periods under the normal ambient. The influence of light exposure on the optical properties of Inq3 thin films were studied by UV–visible absorption, photoluminescence and time-resolved photoluminescence (TRPL) spectroscopic studies. From the photoluminescence decay analysis of as-prepared Inq3 thin films, it is found that the photoluminescence originates from two species having different lifetimes (shorter and longer) corresponding to its two geometrical isomers, namely facial and meridional respectively. The photoluminescence lifetimes are decreasing for increasing light exposure time due to the formation of luminescent quencher during the light exposure. It is reported that the energy of excitons in the light exposed Inq3 thin films can be non-radiatively transferred to its neighbouring luminescent quencher, quenching the photoluminescence of light exposed Inq3 thin films. 相似文献
70.
Sannapaneni Janardan Pothini Suman G. Swapna A. Amrita R. Priya Ramamoorthy Siva Kari Vijayakrishna Akella Sivaramakrishna 《Applied biochemistry and biotechnology》2014,173(2):596-608
A group of hydrazide and semicarbazide derivatives containing isopropylidene, benzylidene, cyclohexylidene, and phospholidene groups was synthesized and characterized by spectroscopic techniques. These compounds were tested for DNA interaction studies monitored by UV-Vis and IR data as well as molecular docking. Investigations on interactions of these compounds with DNA revealed an intercalative mode of binding between them. It is interesting to note that semicarbazide derivatives with aliphatic substituents showed better DNA binding than the aromatic substituents. 相似文献