Sensing cytochrome P450 1A1 activity by a resorufin-based isoform-specific fluorescent probe

Cytochrome P450 1A1 (CYP1A1), a heme-containing monooxygenase, is of particular importance for human health because of its vital roles in the metabolic activation of pro-carcinogenic compounds to the carcinogens. Deciphering the relevance of CYP1A1 to human diseases and screening of CYP1A1 modulators require reliable tool(s) for probing this key enzyme in complex biological matrices. Herein, a practical and ultrasensitive fluorescence-based assay for real-time sensing CYP1A1 activities in biological systems has been developed, via designing an isoform-specific fluorogenic sensor for CYP1A1 (CHPO). The newly developed fluorogenic substrate for CYP1A1 has been carefully investigated in terms of specificity, sensitivity, precision, quantitative linear range and the anti-interference ability. The excellent selectivity, strong anti-interference ability and fast response kinetics, making the practicability of CHPO-based CYP1A1 activity assay is better than that of most reported CYP1A1 activity assays. Furthermore, CHPO has been successfully used for imaging CYP1A1 activities in living cells and human tissues, as well as for high-throughput screening of CYP1A1 inhibitors using tissue preparations as enzyme sources. Collectively, this study provided a practical fluorogenic sensor for real-time sensing CYP1A1 in complex biological systems, which would strongly facilitate the investigations on the relevance of CYP1A1 to human diseases and promote high-throughput screening of CYP1A1 modulators for biomedical applications.     

Li~3Al的几何结构和垂直激发态光谱的量子化学研究

在ab initio DZP水平上, 用能量梯度法对Li~3Al的三种几何构型进行了优化, 并对其中两个能量较低的构型用单、双激发组态相互作用(CISD)进行了垂直跃迁能和振子强度计算, 结果表明: Li~3Al(C~2~v)中存在着三个强度较大的跃迁, 分别是从基态跃迁到1^1B~1, 2^1B~1, 5^1A~1态。Li~3Al(D~3~h)中存在着四个强度较大的跃迁, 分别是从基态跃迁到1^1B~2, 2^1A~1, 3^1B~2, 3^1A~1态。这些强度较大的跃迁均为粒子穴跃迁。     

Li3Al和Li2B2的稳定性和垂直激发态光谱

对于较大的簇合物，电子衍射技术或许能得到有用的信息[1].而小簇合物Mn（3≤n《50）的几何结构就没有标准的方法加以测定．对于二聚体和三聚体[2-5]，振动光谱和转动光谱能够得到精确的核间距和基态的势能面．当n》4时，若要得到一个有价值的结果，需要进行很复杂的振动结构分析．对于这类小簇合物只能得到它们的吸收光谱[6-7].因此，计算其垂直激发态就具有非常重要的意义．用量子化学解释簇合物的吸收光谱已成为一个非常诱人的课题[8].从头算对碱金属簇合物的垂直激发态计算，并与实验光谱进行比较，已成功地预测了一些碱金属簇合物的基…     

红外光谱酰胺Ⅲ带用于蛋白质二级结构的测定研究

用甲醇对BSA和RaseA等蛋白质进行变性处理,结合蛋白质酰胺带的拟合结果对酰胺带各二级结构的谱峰进行了初步指认:1330～1290cm^-1为α-螺旋;1295～1265cm^-1为β-转角;1270～1245cm^-1为无规卷曲;1250～1220cm^-1为β-折叠.依据这些谱峰归属,对一些已知二级结构的蛋白质进行了测定,所得结果与X射线衍射数据以及酰胺带的定量结果基本一致.     

Sensing cytochrome P450 1A1 activity by a resorufin-based isoform-specific fluorescent probe

Cytochrome P450 1A1 (CYP1A1), a heme-containing monooxygenase, is of particular importance for human health because of its vital roles in the metabolic activation of pro-carcinogenic compounds to the carcinogens. Deciphering the relevance of CYP1A1 to human diseases and screening of CYP1A1 modulators require reliable tool(s) for probing this key enzyme in complex biological matrices. Herein, a practical and ultrasensitive fluorescence-based assay for real-time sensing CYP1A1 activities in biological systems has been developed, via designing an isoform-specific fluorogenic sensor for CYP1A1 (CHPO). The newly developed fluorogenic substrate for CYP1A1 has been carefully investigated in terms of specificity, sensitivity, precision, quantitative linear range and the anti-interference ability. The excellent selectivity, strong anti-interference ability and fast response kinetics, making the practicability of CHPO-based CYP1A1 activity assay is better than that of most reported CYP1A1 activity assays. Furthermore, CHPO has been successfully used for imaging CYP1A1 activities in living cells and human tissues, as well as for high-throughput screening of CYP1A1 inhibitors using tissue preparations as enzyme sources. Collectively, this study provided a practical fluorogenic sensor for real-time sensing CYP1A1 in complex biological systems, which would strongly facilitate the investigations on the relevance of CYP1A1 to human diseases and promote high-throughput screening of CYP1A1 modulators for biomedical applications.     

Peptide recognition: encapsulation and alpha-helical folding of a nine-residue peptide within a hydrophobic dimeric capsule of a bowl-shaped host

A dimeric capsule of coordination bowl 1 encapsulated a nine-residue peptide (Trp-Ala-Glu-Ala-Ala-Ala-Glu-Ala-Trp; 2) within the large hydrophobic cavity in water, and stabilized the alpha-helical conformation of bound 2. An NMR titration experiment revealed that monomeric bowl 1 recognized two Trp residues at the both terminals of 2 through 1/2 = 1:1 to 2:1 complexation. The 1:1 and 2:1 species exist in equilibrium even in the presence of excess 1. It was found that the formation of the 2:1 complex, in which two bowls of 1 wrapped the whole of 2, became dominant by the addition of NaNO3 due to the fact that the enhanced ion strength increased the hydrophobic interaction between Trp residues and the cavity of 1. The alpha-helical conformation of 2 within the dimeric capsule of 1 was elucidated from detailed NOESY analysis.     

螯形主体分子的设计、包结性能及其在细辛挥发油 有效成分选择分离中的应用

设计了两种新的具有螯形骨架的主体分子反式-1,2-二苯基-1,2-苊二醇(1)和顺式-1,2-二(1'-萘基)-1,2-苊二醇(2),主体(1),(2)可与许多有机小分子化合物形成配位包合物。用IR和粉末XRD表征了主体分子(1)和(2)的包结物,用^1NMR测定了包结物的主客体分子摩尔比：(1)·DMF(1:2),(1)·DMSO(1:2),(1)·THF(1:2),(1)·二氧六环(1:1),(1)·吡啶(1:1),(2)·DMF(1:1)和(2)·DMSO(1:1)。单晶X射线衍射分析了包结物的晶体结构,(1)·DMF：空间群Pnaa,a=0.9377(1)nm,b=1.4351(1)nm,c=4.0463(3)nm;(1)·DMSO：空间群Pbcn,a=1.6278(1)nm,b=1.0751(1)nm,c=1.4980(1)nm;(2)·DMF：P2~1/n,a=0.9796(1)nm,b=1.2377(1)nm,c=2.2344(3)nm,β=93.02(1)°;游离主体(1)：空间群P1,a=1.0461(1)nm,b=1.1213(1)nm,c=1.5496(1)nm,α=81.74(1)°,β=75.71(1)°,γ=89.00(1)°;分析了主体分子的刚性和柔韧性对包结性能的影响。并研究了主体分子(1)选择分离细辛挥发油,将顺甲基异丁香酚从挥发油中分离出来。     

Synthesis of 2,4-diphenyl-1H-pyrrol-1-amine derivatives

The direction of the reaction of 4-bromo-1,3-diphenyl-2-buten-1-one (γ-bromodypnone) with hydrazines depends on the nature of the substituent they contain. Reaction with 1-methylhydrazinium hydrosulfate gives 1-methyl-3,5-diphenylpyridazin-1-ium bromide but carboxylic acid hydrazides give N-(2,4-diphenyl-1H-pyrrol-1-yl)carboxylic acid amides. γ-Bromodypnone and phenylhydrazine give both 1,3,5-triphenyl-1,4-dihydropyridazine and N,2,4-triphenyl-1H-pyrrol-1-amine (15%). 1-(2,4-Dinitrophenyl)hydrazine gives the 2,4-dinitrophenylhydrazone of (Z)-4-bromo-1,3-diphenyl-2-buten-1-one. Condensation of 2,4-diphenyl-1H-pyrrol-1-amine with aromatic aldehydes readily leads to N-(arylmethylidene)-2,4-diphenyl-1H-pyrrol-1-amines and alkylation with methyl iodide gives N,N-dimethyl-2,4-diphenyl-1H-pyrrol-1-amine. Translated from Khimiya Geterotsiklicheskikh Soedinenii, No. 3, pp. 404–414, March, 2009.     

渤海稠油及其组分中杂原子化合物的负离子电喷雾-高分辨质谱分析

采用沉淀法和色谱分离法将渤海某油田稠油分离成沥青质、胶质、剩余分3个组分。采用负离子电喷雾技术(ESI)结合高分辨傅立叶变换离子回旋共振质谱仪(FT-ICR MS)研究了该稠油及各组分的分子组成。结果表明,剩余分中有较少的极性杂原子化合物能被负离子ESI电离,如N_1,N_1O_1,O_1和O_2类,其等效双键数(DBE)较小。胶质和原油中极性化合物有相同的杂原子类型,包括N_1,N_1O_1,N_1O_2,N_2,N_2O_2S_1,O_1,O_2,其中胶质和原油中N_1,O_1,O_2类化合物的DBE-碳数分布图相似。沥青质中富集高缩合度且多杂原子的酸性化合物,如含杂原子N_,S的氧化程度高的化合物(N_2O_1,N_1O_3,S_1O_3)及O_3类化合物,这些物质具有较高的界面活性,易吸附在界面上促进界面张力降低和增强界面膜强度,从而有利于乳状液稳定存在。N_1O_1,N_1O_2,N_1O_3类化合物,N_2O_1类化合物可能分别是N_1化合物,N_2化合物的氧化降解产物;随着氧化降解程度增加,降解产物的极性明显增强。     

Complete 1H and 13C NMR assignments for two new monodesmoside saponins from Pentaclethra macroloba (Willd.) Kuntze

A detailed NMR study and full assignments of the 1H and 13C spectral data for two novel triterpenoid saponins isolated from the stem bark of Pentaclethra macroloba (Willd.) Kuntze are described. Their structures were established using a combination of 1D and 2D NMR techniques including 1H,1H-COSY, TOCSY, NOESY, gs-HMQC and gs-HMBC, and also electrospray ionization mass spectrometry and chemical methods. The structures were established as 3beta-O-([O-beta-D-glucopyranosyl-(1-->2)-O-beta-D-glucopyranosyl-(1-->4)-O-beta-D-glucopyranosyl-(1-->3)-O-alpha-L-rhamnopyranosyl-(1-->2)]-[O-beta-D-glucopyranosyl-(1-->3)-O-beta-D-glucopyranosyl-(1-->4)])-alpha-L-arabinopyranosylhederagenin (1) and 3beta-O-)[O-beta-D-glucopyranosyl-(1-->2)-O-beta-D-glucopyranosyl-(1-->4)-O-beta-D-glucopyranosyl-(1-->3)-O-alpha-L-rhamnopyranosyl-(1-->2)]-[O-beta-D-glucopyranosyl-(1-->3)-O-beta-D-glucopyranosyl-(1-->4)])-alpha-L-arabinopyranosyloleanolic acid (2).     

Unified Total Synthesis of Polyoxins J,L, and Fluorinated Analogues on the Basis of Decarbonylative Radical Coupling Reactions

Polyoxins J ( 1 a ) and L ( 1 b ) are important nucleoside antibiotics. The complex and densely functionalized dipeptide structures of 1 a and 1 b contain thymine and uracil nucleobases, respectively. Herein we report the unified total synthesis of 1 a , 1 b , and their artificial analogues 1 c and 1 d with trifluorothymine and fluorouracil structures. Decarbonylative radical coupling between α‐alkoxyacyl tellurides and a chiral glyoxylic oxime ether led to chemo‐ and stereoselective construction of the ribonucleoside α‐amino acid structures of 1 a – d without damaging the preinstalled nucleobases. The high applicability of the radical‐based methodology was further demonstrated by preparation of the trihydroxynorvaline moiety of 1 a – d . The two amino acid fragments were connected and elaborated into 1 a – d (longest linear sequence: 11 steps). Compounds 1 a and 1 b assembled in this way exhibited potent activity against true fungi, while only 1 d was active against Gram‐positive bacteria.     

Benchmarking the accuracy of coverage-dependent models: adsorption and desorption of benzene on Pt (1 1 1) and Pt3Sn (1 1 1) from first principles

Bimetallic catalysts have demonstrated properties favorable for upgrading biofuel through catalytic hydrodeoxygenation. However, the design and optimization of such bimetallic catalysts requires the ability to construct accurate, predictive models of these systems. To generate a model that predicts the kinetic behavior of benzene adsorbed on Pt (1 1 1) and a Pt₃Sn (1 1 1) surface alloy (Pt₃Sn (1 1 1)), the adsorption of benzene was studied for a wide range of benzene coverages on both surfaces using density functional theory (DFT) calculations. The adsorption energy of benzene was found to correlate linearly with benzene coverage on Pt (1 1 1) and Pt₃Sn (1 1 1); both surfaces exhibited net repulsive lateral interactions. Through an analysis of the d-band properties of the metal surface, it was determined that the coverage dependence is a consequence of the electronic interactions between benzene and the surface. The linear coverage dependence of the adsorption energy allowed us to quantify the influence of the lateral interactions on the heat of adsorption and temperature programmed desorption (TPD) spectra using a mean-field model. A comparison of our simulated TPD to experiment showed that this mean-field model adequately reproduces the desorption behavior of benzene on Pt (1 1 1) and Pt₃Sn (1 1 1). In particular, the TPD correctly exhibits a broadening desorption peak as the initial coverage of benzene increases on Pt (1 1 1) and a low temperature desorption peak on Pt₃Sn (1 1 1). However, due to the sensitivity of the TPD peak temperature to the desorption energy, precise alignment of experimental and theoretical TPD spectra demands an accurate calculation of the adsorption energy. Therefore, an analysis of the effect of the exchange-correlation functional on TPD modeling is presented. Through this work, we show the necessity of incorporating lateral interactions into theoretical models in order to correctly predict experimental behavior.     

Regio - and stereospecific halogenation of 7alogenoalkenes in liquid hydrogen fluoride

The products of bromo and chlorofluorination of E and Z-1,2-dichloroethylenes, 1, 3-dichloro-1-propenes, 1, 1-dichloro- ethylene and 1, 3-dichloro-2-fluoro-1-propene by N-bromosuccinimide and hexachloromelamine in anhydrous hydrogen fluoride have been studied. It was found that the reaction was in all cases 100% regio and 93–100% trans-stereospecific with the exception of E-1, 2-dichloro-ethylene, its trans-stereospecificity being 85%.Threo and erithro-1-bromo-1, 2-dichloro-2-fluoroethanes, 2-bromo-1, 3-dichloro-1-fluoropropanes and 1, 2, 3-trichloro-1-fluoro-propanes as well 1, 1, 2-trichloro-2-fluoroethane, 1-bromo-2, 2-dichloro-2-fluoroethane, 1, 2, 2-trichloro-2 fluoroethane, 1-bromo-1, 3-dichloro-2, 2-difluoropropane, and 1, 1, 3-trichloro-2,2-difluoropropane were obtained in 50–70% yield.The bromination of E and Z-1, 3-dichloro-1-propenes with molecular bromine in carbon tetrachloride in the dark is non-stereospecific and gives a mixture of erithro and threo-1, 2-dibromo-1, 3-dichloropropanes in the ratio about 1:1. However, the bromination reaction in anhydrous hydrogen fluoride solution proceeds with a high degree of stereospecificity (94–95%) and gives threo-1, 2-dibromo-1, 3-dichloropropane from Z and erithro-1, 2-dibromo-1, 3-dichloropropane from E-1, 3-dichloro-1-propene.The data obtained are considered in terms of an electrophilic mechanism of halogenoalkene halogenation in anhydrous hydrogen fluoride and a free-radical mechanism in carbon tetrachloride.     

Crystal, molecular and electronic structure of N,N'-diphenyl-N,N'-bis(2,4-dimethylphenyl)-(1,1'-biphenyl)-4,4'-diamine and the corresponding radical cation

Oxidation of N,N'-diphenyl-N,N'-bis(3-methylphenyl)-(1,1'-biphenyl)-4,4'-diamine (TPD, 1 a) and N,N'-diphenyl-N,N'-bis(2,4-dimethylphenyl)-(1,1'-biphenyl)-4,4'-diamine (1 b) with SbCl(5) affords the corresponding radical cations quantitatively. The crystal and molecular structure of 1 b and [1 b]SbCl(6), the first tetraphenyl benzidene derivatives to be characterised crystallographically in both the neutral and radical cation states, reveal molecular parameters in agreement with the predictions made on the basis of DFT studies. Analysis of the NIR transition in the radical cations [1](+) (.) allows an estimate of the electronic coupling parameter V (1 a(+) (.) 3200 cm(-1); 1 b(+) (.) 3300 cm(-1)), the reorganisation energy lambda(1 a(+) (.) 7500 cm(-1); 1 b(+) (.) 7800 cm(-1)), and the linear coupling constant l (1 a(+) (.) 3100 cm(-1); 1 b(+) (.) 2700 cm(-1)) of the symmetric mode.     

Electrocatalysis on gold nanostructures: Is the {1 1 0} facet more active than the {1 1 1} facet?

The anisotropic electrocatalytic properties of gold nanobelts and nanoplates enclosed by either {1 1 0} or {1 1 1} facets were studied. Different strategies were used to synthesize these materials. It was found that the {1 1 0} surface of gold does not necessarily show a higher electrocatalytic activity than the {1 1 1} surface. The {1 1 0} surface of gold is more active than the {1 1 1} surface for glucose oxidation in both, neutral and alkaline media. However, for methanol oxidation in alkaline solution, the {1 1 0} surface shows a lower activity than the {1 1 1} surface, which is contrary to the general belief that {1 1 0} facet is the most active surface among the three basal planes. The possible mechanisms are discussed.     

1H-NMR Study on Inclusion Modes of 2-Chloronaphthalene and α-Cyclodextrin in D2

Abstract

With a 1:1 and a 2:1 host-guest stoichiometry, α-cyclodextrin (α-CD) forms inclusion complexes with 2-chloronaphthalene. From simulations concerning observed chemical-shift differences (Δδ_obs) of proton signals of 2-chloronaphthalene, intrinsic Δδ values are estimated for all the guest protons in the 1:1 and 2:1 inclusion complexes. The intrinsic Δδ values indicate that α-CD first binds to a part of a naphthalene ring bearing a C1 atom to form the 1:1 inclusion complex. In the 1:1 and 2:1 inclusion complexes, the symmetry axes of α-CD are tilted approximately 30° relative to a naphthalene longitudinal axis. In the 2:1 inclusion complex, the association through hydrogen bonding most likely occurs between two α-CD molecules whose symmetry axes are laterally shifted.     

Purification of Complex of SAV1 with PP1A

SAV1 is a core component involved in the Hippo pathway which can control the organ size via regulating cell proliferation and apoptosis simultaneously. We explored the regulatory mechanism of SAV1. We established the HEK293T cell pool, the cells in which can stably express SAV1 by retroviruses infection and found that SAV1 stable cells reduced the movement of themselves and resulted in multicellular aggregation. We purified SAV1 interacting protein complex using streptavidin resin and subsequently analyzed the digested peptides by high performance liquid chromatography(HPLC)-MS/MS. Results show that about 150 proteins were identified in the complex of SAV1 with protein. TUBA1A, OTUD4, and ATD were identified as proteins interacting with SAV1. Importantly, PP1A, serine/threonine protein phosphatase PP1-alpha 1 catalytic subunit, was also in the top 10 list. The interaction between PP1A and SAV1 was detected by both co-immunoprecipitation(CO-IP) and immunostaining. Our results indicate that PP1A might be the phosphatase of SAV1 and may take part in the regulation of the Hippo pathway.     

Novel Disulfiram Derivatives as ALDH1a1-Selective Inhibitors

Aldehyde dehydrogenase-1a1 (ALDH1a1), the enzyme responsible for the oxidation of retinal into retinoic acid, represents a key therapeutic target for the treatment of debilitating disorders such as cancer, obesity, and inflammation. Drugs that can inhibit ALDH1a1 include disulfiram, an FDA-approved drug to treat chronic alcoholism. Disulfiram, by carbamylation of the catalytic cysteines, irreversibly inhibits ALDH1a1 and ALDH2. The latter is the isozyme responsible for important physiological processes such as the second stage of alcohol metabolism. Given the fact that ALDH1a1 has a larger substrate tunnel than that in ALDH2, replacing disulfiram ethyl groups with larger motifs will yield selective ALDH1a1 inhibitors. We report herein the synthesis of new inhibitors of ALDH1a1 where (hetero)aromatic rings were introduced into the structure of disulfiram. Most of the developed compounds retained the anti-ALDH1a1 activity of disulfiram; however, they were completely devoid of inhibitory activity against ALDH2.