Oriented nucleation of hydroxylapatite crystals on spider dragline silks

Spider dragline silk as a protein fiber can be pictured as the oriented organization of protein nanocrystals along the long axis with their spacing filled by amorphous protein domains. We used the surface of the spider dragline silk as a biological template to nucleate bone mineral hydroxylapatite (HAP) site-specifically from a HAP-supersaturated solution. HAP crystals were found to be nucleated on the surface of silks with their c axis preferentially oriented at an average angle of 72.9 degrees with respect to the long axis of the silks. The preferred orientation is nearly identical among the different mineralized silks that we studied. Other materials such as Au and CdS could be nucleated on the silks but did not show any preferred orientation. We believe that the oriented nucleation of HAP is directly related to the structures of silks and HAP. The mineralized silks will combine the good mechanical properties of the spider silks and the biocompatibility of HAP and may be assembled into ideal biomaterials as bone implants.     

蜘蛛吐丝过程中钾的作用

用电感耦合等离子体质谱(ICP-MS)对蜘蛛Nephila丝腺体和丝进行测定,结果表明,钾在丝中的含量明显高于在丝腺体中的含量.同时,在蜘蛛丝蛋白溶液中加入氯化钾,溶液出现乳白色浑浊,表明有呈β-折叠构象的微纤产生.浊度测试发现,丝蛋白微纤会逐渐聚集成较大颗粒而在溶液中形成沉淀.另外,红外光谱和拉曼光谱亦证明钾能够使蜘蛛丝蛋白膜发生从无规线团/螺旋到β-折叠的构象转变.有理由认为钾在蜘蛛吐丝过程中起重要作用,它的存在有利于丝蛋白形成β-折叠结构.     

Presence of β-Turn Structure in Recombinant Spider Silk Dissolved in Formic Acid Revealed with NMR

Spider dragline silk is a biopolymer with excellent mechanical properties. The development of recombinant spider silk protein (RSP)-based materials with these properties is desirable. Formic acid (FA) is a spinning solvent for regenerated Bombyx mori silk fiber with excellent mechanical properties. To use FA as a spinning solvent for RSP with the sequence of major ampullate spider silk protein from Araneus diadematus, we determined the conformation of RSP in FA using solution NMR to determine the role of FA as a spinning solvent. We assigned ¹H, ¹³C, and ¹⁵N chemical shifts to 32-residue repetitive sequences, including polyAla and Gly-rich regions of RSP. Chemical shift evaluation revealed that RSP is in mainly random coil conformation with partially type II β-turn structure in the Gly-Pro-Gly-X motifs of the Gly-rich region in FA, which was confirmed by the ¹⁵N NOE data. In addition, formylation at the Ser OH groups occurred in FA. Furthermore, we evaluated the conformation of the as-cast film of RSP dissolved in FA using solid-state NMR and found that β-sheet structure was predominantly formed.     

WISE NMR characterization of nanoscale heterogeneity and mobility in supercontracted Nephila clavipes spider dragline silk

The addition of water to spider dragline silk results in fiber contraction to 50% its initial length and significant changes to the mechanical properties of the silk. This event has been termed supercontraction. A decrease in strength and increase in elasticity have been reported when the silk is in contact with water. Two-dimensional wide-line separation (WISE) nuclear magnetic resonance (NMR) is implemented to correlate (13)C chemical shifts with mobility by observing the corresponding (1)H line widths and line shapes in water-saturated spider dragline silk. The WISE NMR spectrum of the native silk exhibits (1)H line widths that are approximately 40 kHz for all carbon environments characteristic of a rigid organic system. In contrast, the water-saturated case displays a component of the (1)H line that is narrowed to approximately 5 kHz for the glycine C(alpha) and a newly resolved alanine helical environment while the alanine C(beta) corresponding to the beta-sheet conformation remains broad. These results indicate that water permeates the amorphous, glycine-rich matrix and not the crystalline, polyalanine beta-sheets. A delay time is added to the WISE NMR pulse sequence to monitor spin diffusion between the amorphous, mobile region and the crystalline domains. The time required for spin diffusion to reach spatial equilibrium is related to the length scale of the polyalanine crystallites. This technique is employed to measure crystalline domain sizes on the nanometer length scale in water-solvated spider dragline silk. These results provide further insight into the structure of spider silk and mechanism of supercontraction.     

Degree of Biomimicry of Artificial Spider Silk Spinning Assessed by NMR Spectroscopy

Biomimetic spinning of artificial spider silk requires that the terminal domains of designed minispidroins undergo specific structural changes in concert with the β‐sheet conversion of the repetitive region. Herein, we combine solution and solid‐state NMR methods to probe domain‐specific structural changes in the NT2RepCT minispidroin, which allows us to assess the degree of biomimicry of artificial silk spinning. In addition, we show that the structural effects of post‐spinning procedures can be examined. By studying the impact of NT2RepCT fiber drying, we observed a reversible beta‐to‐alpha conversion. We think that this approach will be useful for guiding the optimization of artificial spider silk fibers.     

蚕丝蛋白与类动物丝蛋白聚合物共混膜的振动光谱研究

蚕丝纤维具有优良的力学性能 ,不同的环境条件对其力学性能有一定的影响 ,但其力学性能主要取决于形成纤维过程中所形成的以分子链 β-折叠结构及其沿纤维轴方向高度取向为特征的丝纤维凝聚态结构 [1,2 ] .因此在丝纤维的形成及丝蛋白膜的人工制备过程中 ,丝蛋白分子链的构象及其构象转变一直是研究的重点[3～ 6 ] .以蚕丝蛋白 (Silk Fibroin,SF)稀溶液在常温下浇铸的 SF膜一般以无规线团 /α-螺旋为主的构象状态存在 ;经热处理、极性溶剂 (如甲醇等 )处理、应力作用或共混入一些能与SF形成分子间氢键的聚合物组分后 ,SF膜的构象将从无…     

从天然动物丝到丝蛋白基材料的研究

作为具有优异综合力学性能的天然蛋白质纤维,丰产的动物丝特别是蚕丝长期伴随着人们的日常生活,近十余年来,各种具有特色的功能性丝蛋白基材料更是层出不穷.但在探索动物丝和丝蛋白基材料的过程中,动物丝纤维是经由蚕或蜘蛛等动物的纺器而纺制得到的简单事实往往被忽视;换言之,动物丝实际上是动物对丝蛋白进行体内“加工”后的产物,也是丝蛋白基材料中的一种.因此,天然动物丝中独特的各等级间构效关系与丝蛋白基材料的构效关系之间并不存在着必然的传承效应.本文着重介绍了我们在对动物丝和丝蛋白基材料探索中的经验和体会,即在强调以丝蛋白分子链结构与性能及其之间的关系为研究重点的基础上,从比较和发掘各种天然动物丝的特性入手,进而了解丝蛋白分子链在本体和溶液中的行为,并通过对动物丝蛋白分子链聚集态结构的调控,以达到设计制备一系列多形貌和多功能的动物丝蛋白基材料的目的.     

Observation of spider silk by femtosecond pulse laser second harmonic generation microscopy

An asymmetric β-sheet structure of spider silk is said to induce optical second harmonic generation. In this paper, using an in-house nonscanning type femtosecond pulse laser second harmonic generation microscope, we characterized the behavior of the β-sheet of spider silk under an applied external force. The orientation of the β-sheets was more unidirectional when the silk was extended. One of the origins of the high mechanical strength of the dragline is suggested to be the physical arrangement of its β-sheets.     

Extended wet-spinning can modify spider silk properties

Contrary to expectation, we demonstrate that spider dragline silk spun experimentally under water displays greater stiffness and higher resilience compared to silk spun "naturally" into air. We suggest that this consequence of extended wet-spinning is due to increased molecular orientation resulting from extension of the mobile phase.     

Novel Methods for Obtaining High Modulus Aliphatic Polyamide Fibers

Super high modulus polyethylene fibers can be created by converting high molecular weight flexible PE chains into highly oriented and extended chain conformations. However, unlike polyethylene, aliphatic polyamides have very high cohesive energy and therefore cannot be easily drawn and highly oriented. This review addresses this fundamental problem by analyzing various novel approaches that can be used to suppress hydrogen bonding in these types of polyamides. Plasticization of such polymers with ammonia, iodine, salts, and Lewis acids, as well as dry spinning, wet spinning, and gel spinning, are discussed. Specialized techniques that involve vibrational zone drawing and annealing as well as laser heating zone drawing and annealing are also reviewed. Some of these methods definitely lead to remarkable improvements in initial modulus and other mechanical properties. The development of recombinant spider silk proteins as well progress in spinning these materials is also reported. The advantages and disadvantages of all of these processes are then summarized.     

Determining secondary structure in spider dragline silk by carbon-carbon correlation solid-state NMR spectroscopy

Two-dimensional (2D) (13)C-(13)C NMR correlation spectra were collected on (13)C-enriched dragline silk fibers produced from Nephila clavipes spiders. The 2D NMR spectra were acquired under fast magic-angle spinning (MAS) and dipolar-assisted rotational resonance (DARR) recoupling to enhance magnetization transfer between (13)C spins. Spectra obtained with short (150 ms) recoupling periods were utilized to extract distinct chemical shifts for all carbon resonances of each labeled amino acid in the silk spectra, resulting in a complete resonance assignment. The NMR results presented here permit extraction of the precise chemical shift of the carbonyl environment for each (13)C-labeled amino acid in spider silk for the first time. Spectra collected with longer recoupling periods (1 s) were implemented to detect intermolecular magnetization exchange between neighboring amino acids. This information is used to ascribe NMR resonances to the specific repetitive amino acid motifs prevalent in spider silk proteins. These results indicate that glycine and alanine are both present in two distinct structural environments: a disordered 3(1)-helical conformation and an ordered beta-sheet structure. The former can be ascribed to the Gly-Gly-Ala motif while the latter is assigned to the poly(Ala) and poly(Gly-Ala) domains.     

Correlation between processing conditions,microstructure and mechanical behavior in regenerated silkworm silk fibers

Regenerated silkworm fibers spun through a wet‐spinning process followed by an immersion postspinning drawing step show a work to fracture comparable with that of natural silkworm silk fibers in a wide range of spinning conditions. The mechanical behavior and microstructure of these high performance fibers have been characterized, and compared with those fibers produced through conventional spinning conditions. The comparison reveals that both sets of fibers share a common semicrystalline microstructure, but significant differences are apparent in the amorphous region. Besides, high performance fibers show a ground state and the possibility of tuning their tensile behavior. These properties are characteristic of spider silk and not of natural silkworm silk, despite both regenerated and natural silkworm silk share a common composition different from that of spider silk. © 2011 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys, 2012     

Spinnenseidenproteine: Grundlage für neue Materialien

Spider silk is a biomaterial with extraordinary properties. It is extremely tough and at the same time highly elastic – a combination not found in other polymers. Due to its outstanding potential, spider silk has long been desired as a material for technical applications. This review highlights recent developments in the field of spider silk technology, insights into silk structure, and the natural silk spinning process. Due to the recent progress, spider silk products might be available in the near future, reflecting a new generation of environmentally friendly polymer products.     

Molecular orientation in the Nematic Ordered Cellulose film using polarized FTIR accompanied with a vapor-phase deuteration method

Previously, the authors reported “Nematic Ordered Cellulose (NOC)” that is a well-ordered state of β-1,4-glucan chains without exhibiting typical X-ray diffraction patterns of any cellulose polymorphs (Togawa and Kondo 1999; Kondo et al. 2001; Kondo 2007). The NOC was prepared by stretching water-swollen gel-like films at the draw ratio of 2.0 to provide highly oriented β-1,4-glucan molecular chains of cellulose, which was proved by the high resolution TEM observation. In this paper, a detailed study of the unique ordered state of the NOC was attempted to characterize orientation of the main chains as well as the OH groups of molecules using polarized FTIR accompanied with a vapor-phase deuteration method. The dichroic analysis suggested that the main chains were fairly oriented in the stretching direction whereas the OH groups remained unoriented. The disordered state of the OH groups regardless of the oriented state for the main chain may hinder the oriented crystallization during the preparation of NOC films.     

石墨烯诱导水凝胶成核的高强韧人造蛛丝

天然蜘蛛丝是由β-sheet交联的蛛丝蛋白溶剂流入S-型导管后经牵引拉伸形成,它显示了高强度与高韧性的完美结合。其优异的力学性质主要源于它的多级结构：交联、线性排列的纳米组装体以及核壳结构。受此启发,我们合成了一种交联的水凝胶,通过牵引拉丝的方法,制备了交联的、含有取向排列的纳米组装体结构以及核壳结构的凝胶纤维,并通过少量引入二维纳米材料—氧化石墨烯(0.01%),进一步调控纳米组装体的取向和尺寸,实现了蜘蛛丝般优异的力学性能(断裂强度560 MPa,断裂韧性200 MJ·m^–3,缓冲能94%)。这种纤维可以用于高速下落物体的能量耗散和降低冲击力。     

金属离子导致的丝素蛋白的构象转变

蚕丝和蜘蛛丝的优异力学性能一直是科学家们关注的课题^[1-3]。近年来,在蚕丝蛋白结构及其构象方面的研究取得了许多进展^[3-5]。在蚕的腺体中丝素蛋白的构象为silk I（主要是无规线团为主,还有少量的β－转角,α螺旋等）,而在纤维状的丝中为silk Ⅱ（主要是β折叠）。金属离子在蚕叶丝过程中的作用也一直是一个人们关心的问题。Chen等^[6]在研究丝胶（包附在丝素蛋白表层的另外一种蛋白）时发现,在一定pH条件下,Ni^2 离子通过四配位的螯合作用诱导丝素蛋白β折叠的形成。并且,Viney等^[7]根据电感耦合等离子体（ICP－MS）技术推测Ca^2 的增加能使β折叠的形成加速。     

Research on PAN Nascent Fiber Interior Microstructure through Ultrasonic Etching and Ultrathin Sectioning

The interior microstructures of polyacrylonitrile nascent fibers is studied by the scanning electronic microscopy and the high-resolution transmission electron microscopy through ultrasonic etching and ultrathin sectioning. Due to the orientation and fold of molecular chains, the lamellae of 50–80 nm in thickness are formed. A high number of pores, ranging from dozens to two hundred nanometers in diameters exist between the lamellae, which result from residual solvent. The fibril structure is formed in the nascent fiber during the coagulation process, which are oriented along the fiber axis. An uneven tensile stress distribution leads to the formation of skin-core structures in the nascent fiber during the dry-jet wet spinning process.     

STXM研究氧元素在PAN预氧化纤维截面上的分布

采用扫描透射X射线显微镜( STXM),对聚丙烯腈(PAN)预氧化纤维截面进行具有30 nm空间分辨率的元素分布研究,结合将两幅吸收图上对应像素点的光密度相比求解元素分布的双能衬度分析法,获得氧元素在PAN预氧化纤维截面上的分布信息.STXM实验结果表明,氧元素在PAN原丝截面上分布是均匀的;经过预氧化后,PAN纤维皮...     

Role of Skin Layers on Mechanical Properties and Supercontraction of Spider Dragline Silk Fiber

Spider dragline silk is a composite biopolymer that harbors extraordinary mechanical characteristics, and consists of a hierarchically arranged protein core surrounded by outer “skin” layers. However, the contribution of the successive fiber layers on material properties has not been well defined. Here, the influence of the different components on the physicochemical and mechanical properties of dragline is investigated. The crystal structure and the mechanical properties are not changed significantly after the removal of skin constituents, indicating that the core region of dragline silk fibers determines the structural and mechanical properties. Furthermore, the outer layers have little influence on supercontraction, suggesting they do not function as protection against the penetration of water molecules. On the other hand, the outer layers offer some protection against protease digestion. The present study provides insight into how the function and structure of silk fibers are modulated and facilitates the design of silk‐inspired functional materials.