Plasmodium falciparum RuvB1 is an active DNA helicase and translocates in the 5′–3′ direction

RuvB family of protein contains two similar kinds of proteins i.e. RuvB1 and RuvB2 from yeast to human. These proteins belong to the AAA + class of proteins and are critical components of several multiprotein complexes involved in diverse cellular activities. There are two RuvB proteins annotated in the Plasmodium database but the identification of the third protein recently by our lab has raised the question why Plasmodium falciparum contains three RuvB proteins instead of two. Hence the biochemical characterizations of these proteins have become essential to understand the role of these proteins in the malaria parasite. Recently we have reported the characterization of the recombinant PfRuvB3, which contains ATPase activity but lacks DNA helicase activity. In the present study we report the phylogenetic analysis and detailed biochemical characterization of one of the other RuvB homologue RuvB1 from P. falciparum. PfRuvB1 shows considerable homology with human as well as yeast RuvB1 and contains Walker motif A and Walker motif B. The activity analysis of this protein revealed that PfRuvB1 is an ATPase and this activity increased significantly in the presence of ss-DNA. PfRuvB1 also contains DNA helicase activity and translocates preferentially in 5′ to 3′ direction. In vivo investigation of PfRuvB1 revealed that it is constitutively expressed during all the stages of intraerythrocytic cycle of P. falciparum and localizes mainly to the nucleus. These studies will make important contribution in understanding the role of RuvB protein in P. falciparum.     

A new synthesis and electrochemistry of 1,1′-bis(β-hydroxyethyl)ferrocene

The preparation of 1,1′-bis(β-hydroxyethyl)ferrocene (1) by oxidation of 1,1′-divinylferrocene is described. Compound 1 has been characterized by ¹H and ¹³C{¹H} NMR, and cyclic voltammetry. The electrochemical data are compared to ferrocene and the closely related 2-ferrocenylethanol, 2.     

Synthesis, structure and reactivity of the homoleptic iron(II) complex of the novel 4′-(4?-pyridyl-N-oxide)-2,2′:6′,2″-terpyridine ligand

The new ligand 4′-(4?-pyridyl-N-oxide)-2,2′:6′,2″-terpyridine (pyNoxterpy) and its homoleptic iron(II) complex have been synthesised, and structural and spectroscopic studies have been carried out. The obtained results have been compared with the reported data for the parent ligand 4′-(4?-pyridyl)-2,2′:6′,2″-terpyridine (pyterpy) and its homoleptic iron(II) complex. Significant differences between the spectral and electrochemical properties of the metal complexes have been found, derived from the changes in the electronic properties of the coordinated ligands.     

DNA5′端~(32)P标记方法

在DNA的结构与功能研究中,其5′端~(32)P标记是常用的有效技术。下面我们从《Methods in Enzymolgy》Vol 65中摘译了DNA 5′端脱磷及~(32)P标记方法,供大家参考。 1.用磷酸单酯酶除去DNA链5′末端磷     

Synthesis and characterization of tetrakis(μ-hydroxo)tetrakis(2,2′-dipicolylamine)tetranickel perchlorate, a nickel-hydroxy cubane complex

A novel tetranuclear complex, [Ni(μ₃-OH)(DPA)]₄(ClO₄)₄ (where DPA = 2,2′-dipicolylamine) has been synthesized, with characterization including electronic and infrared spectroscopy, elemental analysis, mass spectrometry, crystal structure analysis, and variable-temperature and variable-field magnetic susceptibility measurements. The complex features a 4Ni-4OH cubane-type cluster, displaying both ferromagnetic and antiferromagnetic intracluster interactions in a 2J model (J₁ = −3.4 cm⁻¹, J₂ = 4.7 cm⁻¹, D = 2.0 cm⁻¹). Each nickel atom sits in a pseudooctahedral environment, with one DPA molecule facially coordinated and the remaining three coordination sites occupied by the bridging hydroxide anions that make up the cubane core.     

Synthesis and characterization of new dinuclear complexes of molybdenum(V) with β′-hydroxy-β-enaminones

New dinuclear molybdenum(V) complexes have been obtained by the reaction of [Mo₂O₃(acac)₄] (acac=acetilacetonate ion) with the polydentate ligands, β′-hydroxy-β-enaminones. All prepared complexes consist of Mo₂O₄ ²⁺ core coordinated by two ligands as in the β-diketonates only through two donor oxygen atoms. Such bonding gives the opportunity for the sixth coordination place around molybdenum to be completed by the monodentate solvent molecule D. All compounds have been characterized by means of elemental analyses, one- and two-dimensional NMR spectroscopy, IR spectroscopy as well as by thermal analyses. The molecular and crystal structures of the molybdenum(V) complexes 1a and 1b coordinated by two different isomeric ligands as well as of the isomer a itself have been determined by a single crystal X-ray diffraction method.     

Synthesis of N-tetra-O-acetyl-β-d-glucopyranosyl-N′-(4′,6′-diarylpyrimidin-2′-yl)thioureas

Some 2-amino-4,6-diarylpyrimidines 2 have been prepared from substituted benzylideneacetophenones and guanidine hydrochloride in the presence of alkali by conventional heating in alcoholic medium and microwave heating in solvent-free conditions. N-(2,3,4,6-Tetra-O-acetyl-β-d-glucopyranosyl)-N′-(4′,6′-diarylpyrimidin-2′-yl)thioureas 4 have been synthesized by reaction of per-O-acetylated glucopyranosyl isothiocyanate 1 and substituted 2-amino-4,6-diarylpyrimidines 2. Two different methods have been used, namely, refluxing in anhydrous dioxane and solvent-free microwave-assisted coupling. The second procedure afforded higher yields in much shorter reaction times. The compounds 2 and 4 were tested for their antibacterial and antifungal activities in vitro against Staphylococcus epidermidis, Enterobacter aerogenes and Candida albicans by disc diffusion method.     

Nucleophilic addition and various species formed in the Cu(II)NCO−3(5)-methylpyrazole system

From the Cu(II)NCO⁻3(5)-methylpyrazole (mpz) system two compounds Cu(NCO)₂(mpz)₂ and four compounds Cu(mpz·NCO)₂ were isolated. The latter compounds contain carbamoylmethylpyrazolate anions as chelate ligands and are coligand isomers of the cyanate compounds. According to the results of indirect structural methods, the Cu(NCO)₂(mpz)₂ complexes have pseudooctahedral structures and differ in their polyhedron distortions. The Cu(mpz·NCO)₂ complexes show tetragonally distorted six or five coordinate structures, possibly differing also by the methyl group position on the pyrazole ring.     

Isomer distribution and structure of (2,2′-biphenyldiolato)bis(β-diketonato)titanium(IV) complexes: A single crystal X-ray, solution NMR and computational study

Novel dichlorobis(β-diketonato)titanium(IV) Ti(C₆H₅COCHCOR)₂Cl₂ and (2,2′-biphenyldiolato)bis(β-diketonato)titanium(IV) Ti(C₆H₅COCHCOR)₂biphen complexes with R = CH₃, C₆H₅ and CF₃, are synthesized and characterized by X-ray crystallography and further physical methods. There is a good agreement between DFT calculated and experimental structural data. A configurational analysis gives a calculated isomer distribution that is in agreement with the experimental data derived from low temperature ¹H NMR spectroscopy. The Ti(C₆H₅COCHCOR)₂biphen complexes exhibit high hydrolytic stability.     

4′-(4-Pyridyl)-2,2′:6′,2″-terpyridine as ligand in the lead(II) complexes, [Pb(pyterpy)(MeOH)I2] · MeOH and [Pb(pyterpy)(μ-AcO)]2(ClO4)2

Two new lead(II) complexes with the ligand 4′-(4-pyridyl)-2,2′:6′,2″-terpyridine (pyterpy), [Pb(pyterpy)(MeOH)I₂] · MeOH and [Pb(pyterpy)(μ-AcO)]₂(ClO₄)₂, have been synthesized and characterized by CHN elemental analysis, ¹H NMR-, ¹³C NMR-, IR spectroscopy and structurally analyzed by X-ray single-crystal diffraction. The thermal stabilities of these compounds were studied by thermal gravimetric (TG) and differential thermal analyses (DTA). The single crystal X-ray analyses show that the coordination number in these complexes is six with three “pyterpy” N-donor atoms and two or three of the anionic ligands. The arrangement of donor atoms in these complexes suggest a gap or hole in the coordination geometry of the lead atoms, possibly occupied by a stereoactive lone pair of electrons on lead(II) and the coordination sphere is hemidirected. The potentially tetradentate ligand 4′-(4-pyridyl)-2,2′:6′,2″-terpyridine (pyterpy) acts as a tridentate donor to Pb(II). The noncoordinated pyridyl group interacts with hydrogen atoms of adjacent molecules and forms normal hydrogen bonds in [Pb(pyterpy)(MeOH)I₂] · MeOH and weak C-H?N interactions for [Pb(pyterpy)(μ-AcO)]₂(ClO₄)₂, thus extending the monomeric structures into one-dimensional networks.     

Synthesis,characterization and study of the PtPt interaction in cis-[(aa)(bb)FCrNCPt(CN)3] (aa,bb=bidentate amine)

Two new dinuclear μ-cyano complexes, cis-[(en)(tn)FCrNCPt(CN)₃] and cis-[(chxn)(tn)FCrNCPt(CN)₃] en=ethylenediamine, tn=1,3-diaminepropane and chxn=1,2-cyclohexanediamine) have been obtained by solid state heating of the trans[Cr(aa)(bb)F(H₂O)][Pt(CN)₄] salts. These complexes have been characterized by chemical analysis, electronic and IR spectra. The dinuclear complexes show strong PtPt interaction both in the solid state and in solution. The association constant of the oligomers formed has been calculated and correlated with the size of the amine ligands. An orbital explanation is proposed to account for the enhancement of the PtPt interaction in the dinuclear complexes relative to the complex salts.     

Specific cation effect on quenching reactions of excited tris(α,α′-diimine) ruthenium(II) and chromium(III) complexes by cyanide complexes in aqueous solutions

Specific salt effects were studied on the quenching reaction of excited [Ru(NN)₃]²⁺ (NN=2,2′-bipyridine(bpy), 1,10-phenanthrorine(phen)) and [Cr(bpy)₃]³⁺ by [Cr(CN)₆]³⁻, [Fe(CN)₆]³⁻ and [Ni(CN)₄]²⁻ in aqueous solutions as a function of alkali metal ions which were added for adjustment of ionic strength. The quenching rate constants in [Ru(NN)₃]²⁺-[Cr(CN)₆]³⁻ and [Cr(bpy)₃]³⁺-[Cr(CN)₆]³⁻ systems are changed by the cations as Li⁺>Na⁺>K⁺≈Rb⁺≈Cs⁺. On the other hand, the rate constants in [Ru(NN)₃]²⁺-[Fe(CN)₆]³⁻ and [Ru(NN)₃]²⁺-[Ni(CN)₄]²⁻ systems, which are diffusion-controlled reactions, are not varied by the alkali metal cations. The obtained order (Li⁺>Na⁺>K⁺≈Rb⁺≈Cs⁺) of the quenching rate constant is quite different from salt effects, Li⁺<Na⁺<K⁺<Rb⁺<Cs⁺, which have been obtained in the electron transfer reactions between complex anions.     

3′-半分子5′-末端双链结构阻止tRNA连接酶酶促反应

用定点突变技术将不同核苷酸引入酵母苯丙氨酸tRNA反密码子环32,37和38位.体外转录制备tRNA前体,其32,37和38位的核苷酸与野生型tRNA前体相应位点的核苷酸不同.用纯化的酵母tRNA内切酶和tRNA连接酶对这些tR-NA前体进行剪接加工.结果说明,这些位点的核苷酸不仅影响tRNA内切酶对tR-NA前体的酶切效率,而且3’-半分子5’-末端双链结构阻止tRNA连接酶将相应的tRNA半分子连接成整分子tRNA.     

Synthesis, properties and crystal structures of new binuclear lead(II) complexes based on phenyl, naphthyl-containing fluorine β-diketones and substituted 2,2′-bipyridines

Four lead(II) complexes with substituted 2,2′-bipyridine adducts and β-diketonates ligands, [Pb(5,5′-dm-2,2′-bpy)(tfpb)₂]₂1, [Pb(4,4′-dmo-2,2′-bpy)(tfpb)₂]₂2, [Pb(4,4′-dm-2,2′-bpy)(tfnb)₂]₂3 and [Pb(5,5′-dm-2,2′-bpy)(tfnb)₂]₂4, (“4,4′-dm-2,2′-bpy”, “5,5′-dm-2,2′-bpy”, “4,4′-dmo-2,2′-bpy”, “Htfpb” and “Htfnb” are the abbreviations of 4,4′-dimethyl-2,2′-bipyridine, 5,5′-dimethyl-2,2′-bipyridine, 4,4′-dimethoxy-2,2′-bipyridine, 4,4,4-trifluoro-1-phenyl-1,3-butanedione and 4,4,4-trifluoro-1-(2-naphthyl)-1,3-butanedione, respectively) have been synthesized and characterized by elemental analysis, IR, ¹H NMR and ¹³C NMR spectroscopy and also studied by thermal and electrochemical as well as X-ray crystallography. The supramolecular features in these complexes are guided/controlled by weak directional intramolecular interactions.     

MonodentateN coordination of 1aza4oxo1,3butadienes [R1NC(R2)C(R3)O] to platinum(II). X-ray structure of trans-[PtCl2(PEt3){σ-N-(t-BuNCHC(Me)O)}]

Reaction of dimeric trans-[PtCl₂(PR₃)]₂ with 1-aza-4-oxo-1,3-butadienes [R¹NC(R²)C(R³)O, R³ = Me, Ph, OMe, NEt₂] in a 1:2 molar ratio results in almost quantitative formation of mononuclear complexes trans·[PtCl₂(PR₃){σ-N-(R¹NC(R²)C(R³)O)}]. The ligands are bonded in the monodentate σ-N bonding mode to the platinum(II) centre. This has been established by an X-ray structure determination of trans-[PtCl₂(PEt₃){σ-N-(t-BuNCHC(Me)O)}]. Crystals of the latter compound are orthorhombic with space group Pc2₁n; cell constants are a = 14.712(3), b = 15.053(2), c = 9.025(5) Å, Z= 4 and R_w = 0.056 for 3281 reflections. The 1aza4oxol,3butadiene (α-iminoketone for R³ is alkyl or aryl) has the E-configuration about the imine bond (CN 1.34(4) Å), with a C(5)C(6) distance of 1.44(5) Å and a NC(5)/ C(6)O torsion angle of 89(4)°. As a result of this ligand conformation, the acetyl hydrogen atoms are positioned (on average) into the neighbourhood of the Pt-atom above the Pt-coordination plane. Infrared and NMR (¹H, ¹³C, ³¹p) data show that these structural features are also predominant in solution.