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151.
Optimized 2DE sample preparation protocols that maximize total protein recovery are fundamental to improving proteome capture and increasing the utility of 2DE, which is in part limited by inadequate recovery of proteins with diverse physicochemical properties. Maintaining protein solubility is an important factor for protein recovery, but the multitude of solubility-enhancing agents and the relatively low-throughput nature of 2DE limit the systematic study of sample preparation. In this work, design of experiment (DOE) approaches are used to optimize protein recovery by altering the levels of four solubility-enhancing agents (urea, DTT, CHAPS, and SDS) in the initial suspension solution. Protein recovery is quantified by a total protein concentration assay, which is demonstrated to be representative of SDS-PAGE and 2DE recovery. DOE methodologies are presented as relatively high-throughput procedures for optimizing 2DE sample preparation parameters for a variety of sample types. Optimal suspension solution compositions are shown to vary across a model protein solution (no urea or DTT), Chinese hamster ovary (CHO) cell lysate (8 M urea, ≥2% CHAPS, ≥32.5 mM DTT), and Escherichia coli cell lysate (8 M urea, 4% CHAPS, 65 mM DTT), with optimized conditions increasing 2DE protein recovery at least 50% compared to suboptimal conditions. 相似文献
152.
Novel X-type polyurethane 5 containing 4-(2′-carbomethoxy-2′-cyano)vinyl-6-nitroresorcinoxy groups as nonlinear optical (NLO) chromophores, which constitute part of the polymer backbone, was prepared and characterized. Polyurethane 5 is soluble in common organic solvents such as acetone and N,N-dimethylformamide. It shows thermal stabilities up to 260°C from thermogravimetric analysis with glass transition temperature obtained from differential scanning calorimetry near 108°C. The second harmonic generation (SHG) coefficient (d33) of poled polymer film at 1064 nm fundamental wavelength is 9.83 × 10?9 esu. Polymer 5 exhibits a high thermal stability even at 2°C higher than Tg, and no significant SHG decay is observed below 110°C, which is acceptable for nonlinear optical device applications. 相似文献
153.
Joong Chul Choe Nu Ri Cheong Seung Min Park 《International journal of mass spectrometry》2009,279(1):25-31
The potential energy surface (PES) for dissociation of aniline ion was determined using density functional theory molecular orbital calculations at the B3LYP/6-311+G(3df,2p)//B3LYP/6-31G(d) level. On the basis of the PES obtained, kinetic analysis was performed by Rice–Ramsperger–Kassel–Marcus (RRKM) calculations. The RRKM dissociation rate constants agreed well with previous experimental data. The most favorable channel was formation of the cyclopentadiene ion by loss of HNC, occurring through consecutive ring opening and re-closure to a five-membered ring. Loss of H could compete with the HNC loss at high energy, which occurred by direct cleavage of an N–H bond or through ring expansion. 相似文献
154.
Jonathan M Choe Deenadayalan Bakthavatsalam Jonathan E Phillips Richard H Gomer 《BMC biochemistry》2009,10(1):1-12
Background
Mycobacterium tuberculosis, an intracellular pathogen encounters redox stress throughout its life inside the host. In order to protect itself from the redox onslaughts of host immune system, M. tuberculosis appears to have developed accessory thioredoxin-like proteins which are represented by ORFs encoding WhiB-like proteins. We have earlier reported that WhiB1/Rv3219 is a thioredoxin like protein of M. tuberculosis and functions as a protein disulfide reductase. Generally thioredoxins have many substrate proteins. The current study aims to identify the substrate protein(s) of M. tuberculosis WhiB1. 相似文献155.
156.
This perspective discusses the use of sequential self-assembly in the construction of metal-organic frameworks through the systematic insertion, replacement, and removal of organic structural building units. We review previous works that can be classified as such sequential self-assembly in multidimensional MOFs. 相似文献
157.
Chung BH Kim S Kim JD Lee JJ Baek YY Jeoung D Lee H Choe J Ha KS Won MH Kwon YG Kim YM 《Experimental & molecular medicine》2012,44(3):191-201
Nitric oxide (NO) produced by endothelial NO synthase (eNOS) plays an important role in vascular functions, including vasorelaxation. We here investigated the pharmacological effect of the natural product syringaresinol on vascular relaxation and eNOS-mediated NO production as well as its underlying biochemical mechanism in endothelial cells. Treatment of aortic rings from wild type, but not eNOS(-/-) mice, with syringaresinol induced endothelium-dependent relaxation, which was abolished by addition of the NOS inhibitor N(G)-monomethyl-L-arginine. Treatment of human endothelial cells and mouse aortic rings with syringaresinol increased NO production, which was correlated with eNOS phosphorylation via the activation of Akt and AMP kinase (AMPK) as well as elevation of intracellular Ca(2+) levels. A phospholipase C (PLC) inhibitor blocked the increases in intracellular Ca(2+) levels, AMPK-dependent eNOS phosphorylation, and NO production, but not Akt activation, in syringaresinol- treated endothelial cells. Syringaresinol-induced AMPK activation was inhibited by co-treatment with PLC inhibitor, Ca(2+) chelator, calmodulin antagonist, and CaMKKβ siRNA. This compound also increased eNOS dimerization, which was inhibited by a PLC inhibitor and a Ca(2+)-chelator. The chemicals that inhibit eNOS phosphorylation and dimerization attenuated vasorelaxation and cGMP production. These results suggest that syringaresinol induces vasorelaxation by enhancing NO production in endothelial cells via two distinct mechanisms, phosphatidylinositol 3-kinase/Akt- and PLC/Ca(2+)/CaMKKβ-dependent eNOS phosphorylation and Ca(2+)-dependent eNOS dimerization. 相似文献
158.
Y0.2CoFe1.8O4 nanopowders were prepared using a sol–gel combustion method. Metal nitrates, such as yttrium nitrate, cobalt nitrate and ferric nitrate, were used as the source materials. Citric acid and polyvinyl alcohol were used as the burning agent and agglomeration reducing agent, respectively. The pH of the precursor was maintained at 7. The mean crystallite size of the prepared ferrite was in the range of ∼20–70 nm. The inverse spinel structure, cubic morphology, and the identification of functional groups of the yttrium-doped cobalt ferrite were analyzed systematically using several analytical tools. 相似文献
159.
Synthesis of highly monodisperse polystyrene microspheres via dispersion polymerization using an amphoteric initiator 总被引:1,自引:0,他引:1
The highly monodisperse polystyrene (PS) microspheres were produced by dispersion polymerization using an amphoteric initiator, 2,2'-azobis [N-(2-carboxyethyl)-2-2-methylpropionamidine] (VA-057). The polymerization characteristics were investigated and compared with conventional initiators, 2,2-azobis(isobutyronitrile) (AIBN) and benzoyl peroxide (BPO). The monodisperse PS microspheres having the coefficient of variation (C(v)) of diameter all less than 4% are obtained at very low stabilizer, poly(vinyl pyrrolidone) (PVP) concentrations of 1 and 2 wt%. It is found that the size dependence of the VA-057 system, D(n) proportional, variant [VA-057](0.267), is less sensitive than a conventional initiator system. When the same amount, 2 wt%, of AIBN, BPO, and VA-057 is used under the identical PVP concentration of 2 wt%, the D(n)/C(v)'s are 1.95/11.57, 1.47/22.44, and 2.08 microm/2.50%, respectively. The uniformity of particles was characterized employing an optical analyzer, Turbiscan. For the VA-057 system, the back scattering intensity is linearly reduced with time having a constant sedimentation rate of 48.98 microm/min throughout the settling process. The uniformity of PS particles in the VA-057 system stems from (1) the higher rate of polymerization in early stage of polymerization, followed by significantly faster reduction of the rate, and (2) good dispersion stability of primary particles. Therefore, it is found that the use of an amphoteric initiator, VA-057, is promising for producing monodisperse particles in dispersion polymerization. 相似文献
160.
Let X be a non-singular complex projective curve of genus ≥3. Choose a point x ∈ X. Let Mx be the moduli space of stable bundles of rank 2 with determinant We prove that the Chow group CHQ1(Mx) of 1-cycles on Mx with rational coefficients is isomorphic to CHQ0(X). By studying the rational curves on Mx, it is not difficult to see that there exits a natural homomorphism CH0(J)→CH1(Mx) where J denotes the Jacobian of X. The crucial point is to show that this homomorphism induces a homomorphism CH0(X)→CH1(Mx), namely, to go from the infinite dimensional object CH0(J) to the finite dimensional object CH0(X). This is proved by relating the degeneration of Hecke curves on Mx to the second term I*2 of Bloch's filtration on CH0(J).
Insong Choe was supported by KOSEF (R01-2003-000-11634-0). 相似文献