Photoactive Ru(II) -polypyridyl complexes that display sequence selectivity and high-affinity binding to duplex DNA through groove binding

The duplex-DNA binding properties of a nonintercalating polypyridyl ruthenium(II) complex that incorporates a linear extended ligand with a catechol moiety has been probed with a variety of photo- and biophysical techniques. These studies reveal that the complex groove binds to DNA sequences biphasically, and displays binding constants equivalent to those of high-affinity metallointercalators. The complex also displays preferential binding to AT-rich sequences. Changes in the structure of the coordinated catechol ligand and the incorporation of intercalating ancillary ligands into the complex were found to modulate both the optical-binding response and binding parameters of the system, which indicates that the catechol moiety plays a crucial role in the observed enhancement to binding affinities.     

Identification of [PtCl2(phen)] binding modes in amyloid-β peptide and the mechanism of aggregation inhibition

Platinum phenanthroline complexes inhibit amyloid-β (Aβ) aggregation and reduce Aβ-caused neurotoxicity [Proc. Natl. Acad. Sci., 2008, 105, 6813-6818]. In this study, we investigated the interactions of Aβ(1-16) with [PtCl(2)(phen)] (phen=1,10-phenanthroline) using HPLC, ESI-MS, and NMR spectroscopy , and characterized the identity of products using tandem mass spectrometry. Results indicated that the phenanthroline ligand could induce noncovalent interactions between Aβ peptide and platinum complexes, leading to rapid Aβ platination. Multiple products were generated in the reaction, in which His6/His14 chelation was preferentially formed. Coordination of Asp7, His13, and Lys16 was also detected in other products. The majority of products were monoplatinated adducts with binding of the {Pt(phen)} scaffold, which impeded intermolecular interactions between Aβ peptides. Moreover, noncovalent interactions were confirmed by the interaction between Aβ peptide and [Pt(phen)(2)]Cl(2). The synergistic roles of the phen ligand and platinum(II) atom in the inhibition of Aβ aggregation are discussed.     

Crystallographic analysis of metal-ion binding to human ubiquitin

The metal-binding ability of human ubiquitin (hUb) towards a selection of biologically relevant metal ions and complexes has been probed. Different techniques have been used to obtain crystals suitable for crystallographic analysis. In the first type of experiments, crystals of hUb have been soaked in solutions containing copper(II) acetate and two metallodrugs, Zeise salt (K[PtCl(3)(η(2)-C(2)H(4))]·H(2)O) and cisplatin (cis-[PtCl(2)(NH(3))(2)]). The Zeise salt is used in a test for hepatitis, whereas cisplatin is one of the most powerful anticancer drugs in clinical use. The Zeise salt readily reacts with hUb crystals to afford an adduct with three platinum residues per protein molecule, Pt(3)-hUb. In contrast, copper(II) acetate and cisplatin were found to be unreactive for contact times up to one hour and to cause degradation of the hUb crystals for longer times. In the second type of experiments, hUb was cocrystallized with a solution of copper(II) or zinc(II) acetate or cisplatin. Zinc(II) acetate gives, at low metal-to-protein molar ratios (8:1), crystals containing one metal ion per three molecules of protein, Zn-hUb(3) (already reported in previous work), whereas at high metal-to-protein ratios (70:1) gives crystals containing three Zn(II) ions per protein molecule, Zn(3)-hUb. In contrast, once again, copper(II) acetate and cisplatin, even at low metal-to-protein ratios, do not give crystalline material. In the soaking experiment, the Zeise anion leads to simultaneous platination of His68, Met1, and Lys6. Present and previous results of cocrystallization experiments performed with Zn(II) and other Group 12 metal ions allow a comprehensive understanding of the metal-ion binding properties of hUb with His68 as the main anchoring site, followed by Met1 and carboxylic groups of Glu16, Glu18, Glu64, Asp21, and Asp32, to be reached. In the case of platinum, Lys6 can also be a binding site. The amount of bound metal ion, with respect to that of the protein, appears to be a relevant parameter influencing crystal packing.     

有机和生物晶体固态核磁共振参数的准确预测

理论计算有助于复杂的有机和生物系统光谱的鉴定.对于核磁共振光谱,固体结晶中的化学位移和四极耦合常数(QCC)受到邻近的分子和晶格的氢键和范德华作用较大的影响,从而显示出与气态单体分子不同的NMR参数.因此,在固体晶体NMR参数的理论计算中有必要将氢键和范德华作用这两个因素考虑进来.基于周期性方法,本文采用L-Ala-Gly二肽和硝基苯晶体作为模型体系来考察该方法计算NMR参数的精度.研究结果显示周期结构模型能够将分子间的氢键和范德华作用考虑进来,得到的化学位移和QCC值明显优于传统的单分子模型和超分子模型得到的结果,采用该方法计算的结果能够重现NMR实验结果.     

一类新型冠醚稀土配合物的合成、表征及与DNA作用研究

合成了四种双(N,N′-乙基,苯基)-2,3-萘-二(氧杂乙酰胺)(L=C30H30N2O4)稀土配合物。通过元素分析、红外光谱、差热-热重及摩尔电导值的分析,确定配合物的组成为[RE(pic)3L],RE=Eu(Ⅲ),Tb(Ⅲ),Ce(Ⅲ),Y(Ⅲ)。电化学实验表明:配体无电化学活性,配合物则呈现出一准可逆的氧化还原过程,在10～1000 mV.s-1扫速范围内配合物的氧化还原峰电流与扫描速度的平方根(ν1/2)呈良好的线性关系,表明其在玻碳电极(GCE)上的伏安行为受扩散控制。另外,通过循环伏安法、光谱法及粘度法研究了配合物与DNA的作用方式,结果表明:配合物均以插入方式与DNA作用,并且作用大小次序是Ce(pic)3L>Tb(pic)3L>Y(pic)3L>Eu(pic)3L。在pH=3.0适宜酸度条件下,建立了以Ce(pic)3L为探针定量测定DNA的分析方法。     

高效亲和色谱法测定2种中药成分与人血清白蛋白的结合率

采用高效亲和色谱技术(HPAC)对中药成分与人血清白蛋白(HSA)的相互作用进行了研究。首先采用点击化学的方法制备了表面键合有HSA蛋白的硅胶固定相并装填成亲和色谱柱,根据药物在该色谱柱上与空白硅胶柱上的保留时间差计算得到药物与蛋白的结合率。利用该方法测得模型化合物华法令与HSA的结合率与文献中采用超滤法测得的结果基本一致,表明该方法可用于测定药物与HSA的结合率。在此基础上用该方法测定了葛根素和告依春两种中药成分与HSA的相对结合率分别为10.26%和10.20%。同时用超滤的方法测定了葛根素与HSA的结合率为14.25%。结果表明,HPAC可以作为研究药物与蛋白相互作用的一种简便可行的方法,其测定结果与超滤方法一致。     

Synthesis, Characterization, DNA-binding Properties and DNA Cleavage of a New Ternary Copper(Ⅱ) Complex with Mixed-ligands of Tridentate Schiff Base and 1,10-Phenanthroline

A new copper(II) complex, [Cu(naph‐leu)phen]CH₃OH·0.5H₂O, in which naph‐leu is the tridentate Schiff base ligand derived from the condensation of 2‐hydroxy‐1‐naphthaldehyde and L‐leucine, phen is phenanthroline, has been synthesized and characterized by elemental analyses, IR spectra and single crystal X‐ray diffraction. The DNA‐binding properties of this complex have been investigated by absorption spectra, fluorescence spectra and circular dichroism (CD) spectra, as well as viscosity measurement. Results show that this copper(II) complex binds to calf thymus DNA (CT‐DNA) in an intercalative mode and its intrinsic binding constant K_b is 4.87×10³ L·mol^?1. Furthermore, the DNA cleavage activity of this copper(II) complex has also been investigated by submarine gel electrophoresis. Interestingly, it was found that this complex can cleave the supercoiled plasmid pBR322 DNA to both nicked and linear forms.     

Crystal structure and DNA binding studies of a cobalt(Ⅱ) complex containing mixed-ligands of 1,10-phenathroline and glycollic acid

A cobalt（11） complex containing mixed-ligands of 1,10-phenathroline（phen） and glycollic acid（GA）.|Co（GA）₂（phen）| was synthesized and its structure was characterized by X-ray diffraction.The interaction of the complex with DNA was investigated by electronic absorption spectra and electrochemical methods.Electronic absorption spectrum experiments showed that after interaction with DNA.the characteristic absorption peaks of |Co（GA）₂（phen）| underwent hypochromic effect as well as redshift. Also,the binding strength of 3.8×10⁴L/mol was estimated by titration method.Electrochemical assays revealed that the redox peak currents of the complex decreased obviously accompanied by a positive shift of the formal potential after association with DNA.All these results revealed that the synthesized cobalt complex bound with DNA via an intercalation mode.     

Biophysical studies of the interaction between a triazole derivative and bovine serum albumin by multi-spectroscopic and molecular modeling methods

The interaction between 3-thiol-4-(2,4-dichlorobenzylideneamino)-5-methyl-4H-1,2,4-triazole (CBTZ) and bovine serum albumin (BSA) under physiological conditions was investigated by fluorescence,UV-vis absorption and circular dichroism (CD) spectroscopy as well as molecular modeling methods. The result of fluorescence experiment indicates the static quenching as a result of the formation of the CBTZ-BSA complex. The binding constants (Ka) at different temperatures were calculated according to the modified St...