Self-assembly of a hydrophobic polypeptide containing a short hydrophilic middle segment: Vesicles to large compound micelles

This report describes a facile route to prepare the vesicles and large compound micelles (LCMs) from a series of poly(epsilon-benzyloxycarbonyl L-lysine)-block-poly[diethylene glycol bis(3-amino propyl) ether]-block-poly(epsilon-benzyloxycarbonyl L-lySine) (PZLL-DGBE-PZLL) in their water solution, depending on molecular weight of the polypeptides. A pyrene probe is used to demonstrate the aggregate formation of PZLL-DGBE-PZLL in solution, and also to measure their critical micelle concentration as a function of molecular weight of the polymer.

Hollow DNA/PLL microcapsules with tunable degradation property as efficient dual drug delivery vehicles by alpha-chymotrypsin degradation

Hollow deoxyribonucleic acid (DNA)/poly-L-lysine (PLL) capsules were successfully fabricated through a layer-by-layer (LbL) self-assembly of DNA and PLL on porous CaCO3 microparticles, followed by removal of templates with ethylenediamine tetraacetic acid disodium salt (EDTA). The enzymatic degradation of the capsules in the presence of alpha-chymotrypsin was explored. The higher the enzyme concentration, the higher is the degradation rate of hollow capsules. in addition, glutaric dialdehyde (GA) cross-linking was found to be another way to manipulate degradation rate of hollow capsules.

A facile pathway to prepare enzymatically degradable microcapsules with tunable capsule shell properties

Dextran sulfate (DS)/poly-L-lysine (PLL) microcapsules are fabricated by an in situ coacervation method using DS-doped CaCO3 microparticles as templates. Twinned superstructures or spherical CaCO3 microparticles are produced depending on DS concentration in the starting Solution. DS/PLL microcapsules with ellipsoidal or spherical outline are obtained after removal of templates in disodium ethylene diamine tetraacetate dehydrate (EDTA) without PLL. Their shell thickness and negative surface charges increase with the DS weight percentage in the templates. The surface potential of DS/PLL microcapsules.

Self-Assembly of Hyperbranched Multiarmed PEG-PEI-PLys(Z) Copolymer into Micelles, Rings, and Vesicles

Self-assembling of synthesized novel biodegradable hyperbranched amphiphilic poly(ethylene glycol)-polyethylenimine-poly(epsilon-benzyloxycarbonyl-L-lysine) (PEG-PEI-PLys(Z)) in aqueous media is studied. In aqueous media. PLys(Z) is the hydrophobic segment, with PEG and PEI as the hydrophilic segments. It will self-assemble into spherical shape when the selected solvent water is dropped into the common solvent tetrahydrofuran (THF). And when PEG-PEI-PLYS in common solvent is dropped into mixed solvent water and THF, rings will come into King. The spherical and rings are observed by environmental scanning electron microscopy (ESEM) and transmission electron microscopy ITEM). It shows that the size of the sphere is about 100 nm, and the diameter of ring distributes from 400 nm to 10 mu m and bigger with the time roll around.

Facile coprecipitation to synthesize water-soluble amine-capped magnetite nanoparticles

Water solubility and surface functionalization of magnetic nanoparticles are crucial for bioapplication.[1]In this study,we presented a facile coprecipitation approach to synthesize lysine stabilized Fe3O4 nanoparticles.Lysine functionalized magnetite nanoparticles show an excellent colloidal stability of ＞20h.The as-synthesized magnetite nanoparticles have abundant amine groups on their surface which provide convenient sites for covalent linking of biological macromolecules.We believe that these amine-functionalized magnetic nanoparticles can be potentially used in fields such as magnetic bio-separation,immunoassay,MRI,and targeted drug delivery.

复方板蓝根颗粒水提部位的ESI-MS指纹图谱研究

Sialic acid and lysine as an internal standard substance,the ESI-MS fingerprinting of extracts of Compound Indigowoad Root Granule which from different origin was studied.The fingerprinting was obtained with better precision and reproducibility.This work can provide a reference for the quality control of Compound Indigowoad Root Granule.

Direct formation of giant vesicles from synthetic polypeptides

This report describes direct formation of giant vesicles from a series of poly(L-lysine)-block-poly(L-phenylalanine) (PLL-b-PPA) block copolymers from their water solution. These polymers are prepared by successive ring-opening polymerization (ROP) of the two alpha-amino acid N-carboxyanhydrides and then removing the side chain protecting groups by acidolysis. The structures of the copolymers are confirmed by nuclear magnetic resonance (NMR), differential scanning calorimetry (DSC), and size exclusion chromatography ( SEC). The vesicles are studied by atomic force microscopy (AFM), field emission scanning electron microscopy (ESEM), and confocal laser scanning microscopy (CLSM). Rhodamine B is used as a fluorescent probe to confirm the existence of the vesicle with an aqueous interior. The vesicle size is in the range 0.55-6 mu m, depending on the absolute and relative lengths of the two blocks, on initial polymer concentration, and on solution pH. The vesicles are still stable in water for 2 months after preparation. Addition of the copolymer to DNA solution results in complex formation with it. The complex assumes the morphology of irregular particles of less than 2 mu m. It is expected to be used in drug and gene delivery.

Synthesis of a novel structural triblock copolymer of poly(gamma-benzyl-L-glutamic acid)-b-poly(ethylene oxide)-b-poly(epsilon-caprolactone)

A novel structural triblock copolymer of poly(gamma-benzyl-L-glutamic acid)-b-poly(ethylene oxide)-b-poly(epsilon-caprolactone) (PBLG-PEO-PCL) was synthesized by a new approach in the following three steps: (1) sequential anionic ring opening polymerization (ROP) of ethylene oxide and epsilon-caprolactone with an acetonitrile/potassium naphthalene initiator system to obtain a diblock copolymer CN-PEO-PCL with a cyano end-group; (2) conversion of the CN end-group into NH2 end-group by hydrogenation to obtain NH2-PEO-PCL; (3) ROP of gamma-benzyl-L-glutamate-N-carboxyanhydrides (Bz-L-GluNCA) with NH2-PEO-PCL as macroinitiator to obtain the target triblock copolymer. The structures from CN-PEO precursor to the triblock copolymers were confirmed by FT-IR and H-1 NMR spectroscopy, and their molecular weights were measured by gel permeation chromatography. The monomer of Bz-L-GluNCA can react almost quantitatively with the amino end-groups of NH2-PEO-PCL macroinitiator by ROP.

Partitioning behavior of amino acids in aqueous two-phase systems containing polyethylene glycol and phosphate buffer

The partitioning behavior of four amino acids, cysteine, phenylalanine, methionine, and lysine in 15 aqueous two-phase systems (ATPSs) with different polyethylene glycol (PEG) molecular weights and phosphate buffers has been studied in the present paper. The phase diagrams of the systems are investigated together with the effect of the PEG molecular weight and pH of the phosphate solutions. The composition of these systems and some parameters such as density and refractive index are determined. The influences of salts in ATPSs, side chain structure of the amino acids, pH of ATPSs, and the PEG molecular weight on the distribution ratios of the amino acids have been studied. This work is useful for the purification of amino acids and the separation of some proteins whose main surface exposed amino acid residues are these four amino acids, respectively.

Biodegradable amphiphilic triblock copolymer bearing pendant glucose residues: Preparation and specific interaction with Concanavalin A molecules

A novel biodegradable amphiphilic block copolymer PLGG-PEG-PLGG bearing pendant glucose residues is successfully prepared by the coupling reaction of 3-(2-aminoethylthio) propyl-R-D-glucopyranoside with the pendant carboxyl groups of PLGG-PEG-PLGG in the presence of N,N'-carbonyldiimidazole. The polymer PLGG-PEG-PLGG, i.e., poly {(lactic acid)-co-[(glycolic acid)-alt-(L-glutamic acid)]}-block-poly(ethylene glycol)-block-poly{( lactic acid)-co-[( glycolic acid)-alt-(L-glutamic acid)]}, is prepared by ring-opening copolymerization of L-lactide (LLA) with (3s)-benzoxylcarbonylethylmorpholine-2,5-dione (BEMD) in the presence of dihydroxyl PEG with molecular weight of 2000 as macroinitiator and Sn(Oct)(2) as catalyst, and then by catalytic hydrogenation. The glucose-grafted copolymer shows a lower degree of cytotoxicity to ECV-304 cells and improved specific recognition and binding with Concanavalin A (Con A). Therefore, this kind of glucose-grafted copolymer may find biomedical applications.

Photoreduction behaviors of microperoxidase-11 adsorbed on the roughened silver electrodes

It was found that microperoxidase-ll (MP-II) can undergo photoreduction at the bale roughened silver electrode. No photoreduction happens at the roughened silver electrode modified with mercaptoundecanoic carboxylic acid/poly-lysine. The photoreduction mechanism is discussed.

The suppression of Ge-132 on the Maillard reaction of BSA by fluorescence spectrum

The fluorescences of BSA and glycosylated BSA were observed respectively. The lambda(cm) of BSA was 340 nm; while the lambda(cm) of glycosylated BSA was 436 nm. Because the fluorescence spectra of them were different greatly, we can observe the suppression of Ge-132 on the Maillard reaction of BSA without any interference of itself. It was showed that the fluorescence intensity of glycosylated BSA increased continuously with the cultured time, Ge-132 may suppress the Maillard reaction of BSA greatly, and the suppressing efficicency would be 32 %. The key site of the Maillard reaction of BSA is free amino groups of alanine residues on N-terminal amino group, besides the epsilon-amino groups of intrachain Lysine residues.

Screening of genes related to ovary development in Chinese shrimp Fenneropenaeus chinensis by suppression subtractive hybridization

The ovary of triploid shrimp Fenneropenaeus chinensis was apparently impaired compared to that of the diploid shrimp at the same age. Therefore triploid shrimp ovary is possible to be taken as a model to understand the mechanism of ovary development of shrimp compared to that of the ovary of diploid shrimp at the same age. In the present study, a suppression subtractive hybridization (SSH) technique was applied to identify differentially expressed genes in the ovary between diploid and triploid shrimp. For the forward library (RNA from the ovary of triploid shrimp as the tester), 54 genes were identified. For the reverse library (RNA from the ovary of diploid shrimp as the tester), 16 genes were identified. The identified genes encoded proteins with multiple functions, including extracellular matrix components, cytoskeleton, cell growth and death, metabolism, genetic information processing, signal transduction/transport or immunity related proteins. Eleven differentially expressed genes were selected to be confirmed in the ovaries of triploid and diploid shrimp by semi-quantitative RT-PCR. Genes encoding spermatogonial stem-cell renewal factor, cytochrome c oxidase subunits I and II, clottable protein, antimicrobial peptide and transposase showed up-regulated expressions in the ovary of triploid shrimp. Genes encoding tubulin, cellular apoptosis susceptibility protein, farnesoic acid O-methyltransferase, thrombospondin and heat shock protein 90 genes showed higher expressions in the ovary of diploid shrimp. The differential expressions of the above genes are suggested to be related to the ovary development of shrimp. It will provide a new clue to uncover the molecular mechanisms underlying the ovarian development in penaeid shrimp. (C) 2010 Elsevier Inc. All rights reserved.

栉孔扇贝核心组蛋白的基因结构及H2A抗菌活性的研究

栉孔扇贝是我国传统的海水养殖品种，但自1997 年以来，养殖扇贝陆续爆发的大规模死亡，不但造成了巨大的经济损失，而且严重影响了该产业的健康发展。目前，虽然针对扇贝养殖环境、病原以及养殖技术等方面开展了大量的研究工作，提出了许多防病治病的措施，并取得了一定的成效。但由于引起养殖扇贝病害的病原和发病原因的多样性，大量使用抗菌素和农药后造成病原微生物抗药性的提高以及对环境造成的严重破坏，贝类养殖业要摆脱病害的困扰，必须开辟新的疾病防治途径。 从扇贝自身的免疫防御因子入手，筛选和克隆参与免疫防御的功能基因，尤其是一些新颖的具有抗菌活性的分子，对于深入探讨扇贝的免疫防御机制，指导扇贝的遗传改良和抗病品系的培育具有重要的意义；另一方面，可对抗菌效应物实现重组表达，开发新型的病害预防治疗制剂，取代目前普遍使用的抗生素和化学药物。抗菌效应物是机体在免疫应答过程中产生的多肽类物质，对侵入生物体内的细菌、病毒具有很强的免疫杀灭作用，对抗菌效应物的研究有助于深入了解机体先天性免疫防御的机制。 本研究在同源克隆策略的基础上，从利用构建的Genome Walking 文库中克隆到了栉孔扇贝核心组蛋白群的全长序列，该串联重复序列全长5671bp，包括各一个拷贝的组蛋白H4, H2B, H2A 和 H3。所有的核心组蛋白在3’侧翼序列均具有与其在细胞周期进化模式相关的特征结构，即两个不同的终止信号：发卡结构和至少一个多聚腺苷酸信号序列（AATAAA）。在5’区域的起始密码子上游37–45 bp处的保守的CAP位点（5’-PyCATTCPu-3’）存在于除H2B外的每一个基因中；规则的TATA 和CAAT元件也在核心组蛋白群中的个别的基因中找到。在H2B 和H2A基因的启动子区域，对于定位转录起始位点非常重要的元件（5’-GATCC-3’）也相对保守； 在H2B启动子区域存在着与其特征序列（5’-GGAATAAACGTATTC-3’）相似性很高的序列结构5’-GGATCGAAACGTTC-3’。增强子序列只发现存在于H4 和 H3基因中，其序列结构与组蛋白增强子序列(5’-TGATATATG-3’)基本匹配。在组蛋白基因群中存在着一些保守的序列和重复结构表明组蛋白基因的进化是采取“生与死的进化模 式”并伴随着强的纯化选择压力，使得该基因群变异较少以保持其基本功能。同时，利用18S rRNA做参照，探讨了H2A 和H2B作为分子系统进化分析的潜在分子标记，表明组蛋白H2A 和H2B可以作为分子系统进化分析得候选分子，它们在区分近缘种的分辨率上表现出了更高的灵敏度。该研究结果为进一步定性软体动物组蛋白重复单位提供了基础。 在脊椎动物中，组蛋白H2A通过特异性剪切其N末端产生新颖的抗菌肽的形式来参与宿主的免疫应答反应，在软体动物中是否存在同样的机制还未有研究报道。本研究利用上述克隆的H2A基因研究了其在病原胁迫下的表达变化规律并对其N末端39aa进行了重组表达和抗菌活性分析，以期为开发和利用软体动物的新颖的抗菌肽提供理论依据。半定量RT-PCR发现血细胞中H2A 的mRNA 在微生物感染前后的表达量没有任何显著的变化，表明H2A本身并不直接参与对病原的清除过程或者说病原微生物并不能诱导H2A的表达。因此，我们推测该基因可能象脊椎动物一样以前体形式存在，经剪切后参与宿主的免疫应答过程，为此我们研究了H2A的N末端的抗菌活性。通过将与脊椎动物buforin I同源的H2A的N末端39aa克隆到毕赤酵母表达载体pPIC9K实现了该基因N末端的重组表达。抑菌实验表明，重组产物具有广谱的抗菌活性，其对供试的革兰氏阳性菌藤黄微球菌表现出显著的抗菌活性，而对革兰氏阴性菌（鳗弧菌、亮弧菌）的抑菌活性则相对较弱；此外，重组产物对毕赤酵母GS115也表现出一定的杀菌活性，证明其具有抗真菌活性。上述研究结果证明组蛋白H2A的N末端是一种潜在的抗菌肽，但该抗菌肽是否参与机体的免疫应答过程需要进一步的深入研究。

Bromophenol derivatives from the red alga Rhodomela confervoides

Eight new bromophenol derivatives, 2,3-dibromo-4,5-dihydroxybenzyl methyl sulfoxide (1), 4-(2,3-dibromo-4,5-dihydroxyphenyl)-3-butene-2-one (2), 2-(3-bromo-5-hydroxy-4-methoxyphenyl)-3-(2,3-dibromo-4,5-dihydroxyphenyl)propionic acid (3), 2-(3-bromo-5-hydroxy-4-methoxyphenyl)-3-(2,3-dibromo-4,5-dihydroxyphenyl)propionic acid methyl ester (4), 2-phenyl-3-(2,3-dibromo-4,5-dihydroxyphenyl)propionic acid (5), 4'-methoxy-2",3',3"-tribromo-4",5',5"-trihydroxydiphenylacetic acid (6), and 3-bromo-5-hydroxy-4-methoxyphenylacetic acid (7) and its methyl ester (8), together with a known bromophenol, 3-bromo-5-hydroxy4-methoxybenzoic acid (9), were isolated from the red alga Rhodomela confervoides. Their structures were elucidated by spectroscopic methods including IR, EIMS, FABMS, ESIMS, HRFABMS, HRESIMS, 1D and 2D NMR, and single-crystal X-ray structure analysis. Compounds 1-4, 8, and 9 were found inactive against several human cancer cell lines and microorganisms.