Kinetic analysis of cellular internalization and expulsion of unstructured D-chirality cell penetrating peptides

Most cell penetrating peptides (CPPs) are unstructured and susceptible to proteolytic degradation. One alternative is to incorporate D-chirality amino acids into unstructured CPPs to allow for enhanced uptake and intracellular stability. This work investigates CPP internalization using a series of time, concentration, temperature, and energy dependent studies, resulting in a three-fold increase in uptake and 50-fold increase in stability of D-chirality peptides over L-chirality counterparts. CPP internalization occurred via a combination of direct penetration and endocytosis, with a percentage of internalized CPP expelling from cells in a time-dependent manner. Mechanistic studies identified that cells exported the intact internalized D-chirality CPPs via an exocytosis independent pathway, analogous to a direct penetration method out of the cells. These findings highlight the potential of a D-chirality CPP as bio-vector in therapeutic and biosensing applications, but also identify a new expulsion method suggesting a relationship between uptake kinetics, intracellular stability, and export kinetics.     

Effect of amide type modified rapeseed oil as lubricating additive on friction and wear behavior of steel-steel and steel-aluminum alloy systems

A new type of environmentally friendly lube additive-amide type modified rapeseed oil was synthesized and characterized by infrared spectrum. Its effect on the friction and wear behavior of steel-steel and steel-aluminum alloy systems were investigated with a four-ball machine and an Optimol SRV friction and wear tester respectively.The morphographies of the worn surfaces were analyzed by means of scanning electron microscopy(SEM). The worn surfaces of the 2024Al alloy block were analyzed by means of X-ray photoelectron spectroscopy(XPS). The results show that the modified rapeseed oil as additives can obviously decrease the wear rate and friction coefficient of steel pair and steel-aluminum frictional pair. Its lubrication mechanism is inferred that a high strength complex protection films form on the worn surface of the Al alloy due to the adsorption or tribochemistry reaction of a long chain additive molecule and high reaction activity of N element.     

Electrostatic force microscopy of self-assembled peptide structures

In this report electrostatic force microscopy (EFM) is used to study different peptide self-assembled structures such as tubes and particles. It is shown that not only geometrical information can be obtained using EFM, but also information about the composition of different structures. In particular we use EFM to investigate the structures of diphenylalanine peptide tubes, particles, and CSGAITIG peptide particles placed on pre-fabricated SiO(2) surfaces with a backgate. We show that the cavity in the peptide tubes could be due to the presence of water residues. Additionally we show that self-assembled amyloid peptides form spherical solid structures containing the same self-assembled peptide in its interior. In both cases transmission electron microscopy is used to verify these structures. Further, the limitations of the EFM technique are discussed, especially when the observed structures become small compared with the radius of the AFM tip used. Finally, an agreement between the detected signal and the structure of the hollow peptide tubes is demonstrated.     

Peptide Nanophotonics: From Optical Waveguiding to Precise Medicine and Multifunctional Biochips

Optical waveguiding phenomena found in bioinspired chemically synthesized peptide nanostructures are a new paradigm which can revolutionize emerging fields of precise medicine and health monitoring. A unique combination of their intrinsic biocompatibility with remarkable multifunctional optical properties and developed nanotechnology of large peptide wafers makes them highly promising for new biomedical light therapy tools and implantable optical biochips. This Review highlights a new field of peptide nanophotonics. It covers peptide nanotechnology and the fabrication process of peptide integrated optical circuits, basic studies of linear and nonlinear optical phenomena in biological and bioinspired nanostructures, and their passive and active optical waveguiding. It is shown that the optical properties of this generation of bio‐optical materials are governed by fundamental biological processes. Refolding the peptide secondary structure is followed by wideband optical absorption and visible tunable fluorescence. In peptide optical waveguides, such a bio‐optical effect leads to switching from passive waveguiding mode in native α‐helical phase to an active one in the β‐sheet phase. The found active waveguiding effect in β‐sheet fiber structures below optical diffraction limit opens an avenue for the future development of new bionanophotonics in ultrathin peptide/protein fibrillar structures toward advanced biomedical nanotechnology.     

基于RGD多肽掺杂聚吡咯-铟锡氧化物的仿生微电极构建及用于细胞生物学行为的电化学阻抗谱检测

构建一种基于RGD多肽分子掺杂聚吡咯膜修饰的铟锡氧化物微电极(PPy/RGD-ITO),并以此作为传感电极实现细胞生物学行为的电化学阻抗谱检测.采用光刻技术蚀刻感光干膜绝缘层制备ITO微电极;以含RGD模体的多肽分子作为吡咯电聚合唯一的掺杂阴离子,通过电化学共聚合方式在ITO微电极表面沉积PPy/RGD复合膜形成PPy/RGD-ITO微电极;原子力显微镜(AFM)、接触角测量仪和傅里叶变换红外光谱仪(FTIR)分别表征PPy/RGD复合膜的表面拓扑形貌、湿润性和组成成分;人肺癌细胞株A549铺展、粘附及增殖实验考察了PPy/RGD复合膜与细胞间的相互作用;以构建的PPy/RGD-ITO微电极作为传感电极,通过电化学阻抗谱技术对A549细胞粘附增殖行为及天然抗癌药物分子重楼皂苷I的细胞毒性进行了分析.结果显示,通过简单的电化学共聚合成功将RGD分子掺杂进PPy膜内,且PPy/RGD复合膜具有优异的表面物理性能;PPy基质膜内掺杂的RGD分子保留其生物活性,相比裸ITO电极和聚4-苯乙烯磺酸钠(PSS)掺杂的PPy膜,PPy/RGD复合膜能更好地促进A549细胞的铺展、粘附和增殖;由于PPy/RGD-ITO微电极表面A549细胞形态学变化可改变电极系统的阻抗谱特征,因此通过电化学阻抗谱技术可解析A549细胞粘附增殖行为学信息,同时可定量分析重楼皂苷I细胞毒性.因此,通过简单的电化学共聚合方法将生物活性RGD分子掺杂进PPy膜内制备出的PPy/RGD膜具有优良的生物相容性,可作为一种重要的仿生电极修饰材料用于构建电子系统和细胞生物学系统的耦合界面,未来可应用于细胞生物学行为及药物筛选研究.     

类弹性蛋白多肽的分子动力学研究

类弹性蛋白多肽因其具有特殊的相变性质,故而在重组蛋白纯化方面展现出良好的应用前景,对其发生相变的机理进行研究具有重要意义.本文利用同源建模的方法构建了类弹性蛋白多肽的三维结构并进行能量优化,之后采用分子动力学模拟手段,在300 K～400 K间5个不同温度下,对含有100个氨基酸残基的类弹性蛋白多肽[KV8F-20]各进行了6 ns的模拟.模拟过程中,类弹性蛋白多肽发生疏水缩聚,初始结构变得更加紧凑,且温度越高折叠程度越大.水分子在类弹性蛋白多肽的相变行为中起到关键作用.经分析,结果发现疏水作用与水的排出在类弹性蛋白多肽发生相变过程中起到关键作用,类弹性蛋白多肽随着温度的升高,在疏水作用驱动下,其构象折叠程度因疏水缩聚而变得更大.此外,比较不同温度下蛋白的缩聚程度,推断类弹性蛋白多肽在375 K左右发生相变,这与实验观测的结果基本吻合.据此,推测该类弹性蛋白多肽变温度范围约为95℃～102℃.这对后续调控类弹性蛋白多肽的相变具有指导作用.     

枯草芽孢杆菌谷氨酰胺转氨酶的异源表达

以野生型枯草芽孢杆菌基因组DNA为模板,PCR扩增获得带有sD序列及谷氨酸棒状杆菌信号肽AS0949的BTG基因。将其与大肠杆菌-谷氨酸棒状杆菌穿梭表达载体pXMJ19连接,构建重组质粒pXMJ19-Sbtf转化谷氨酸棒状杆菌ATCC13032。经IPTG诱导后该重组茵发酵液具有交联酪蛋白的能力,表明该重组茵能够实现分泌表达。     

Raney-Ni催化下菜籽油生物柴油催化转移加氢的研究

在水环境下,以异丙醇为供氢体和Raney-Ni为催化剂,对菜籽油经酯交换得到的生物柴油进行催化转移加氢研究.通过定时取样分析生物柴油的碘值变化和组分含量,研究了生物柴油中亚麻酸甲酯(C18∶3)、亚油酸甲酯(C18∶2)和油酸甲酯(C18∶1)催化转移加氢选择性和反应动力学,以及加氢过程中不饱和脂肪酸甲酯的顺反异构情况.结果表明:在水环境下生物柴油催化转移加氢反应可用拟一级动力学模型来描述;C18∶3和C18∶2的加氢选择性较高,反应速率常数是C18∶1的6倍,但产物主要为反式油酸甲酯(t-C18∶1),而且顺式油酸甲酯(c-C18∶1)进一步加氢变成硬脂酸甲酯(C18∶0)的速率远远高于t-C18∶1.