小远志多糖一级结构的初步研究

从药用植物小远志中提取、精制得多糖PP-1,PP-2,采用纸层析、薄层层析和气相色谱等方法确定了多糖中的单糖组成.通过高碘酸氧化、Smith降解和甲基化分析等方法确定了相邻单糖基连接方式,初步推测了两个小远志多糖可能的一级结构,为进一步合成和研究其生物活性提供了数据支持.     

沙冬青抗冻蛋白热滞活性的DSC研究

在某些极地鱼类的血清以及某些昆虫和植物中 ,人们发现了一类特殊的物质 .它们的共同特点是能通过直接与冰晶核相互作用,抑制冰晶核的形成和生长,从而降低溶液冰点 .现已发现的这类抗冻物质都属于蛋白质,被统称为抗冻蛋白（ Antifreeze Proteins, AFPs） .抗冻蛋白能以非依数性形式大大降低溶液的冰点,但对熔点的影响很微弱,而且遵从依数性的原则,使冰点低于平衡态的熔点,溶液处于反常的非平衡相变状态 .这种冰点低于熔点的特性称为热滞 (Hysteresis).因此,抗冻蛋白也叫热滞蛋白或温度迟滞蛋白（ Thermal Hysteresis Proteins, …     

猪精浆中附睾特异性蛋白质的纯化及其一级结构的研究

哺乳动物体中附睾内精子的成熟过程要经过配子融合而形成合子的过程,其中,精子聚合到附睾分泌的特异蛋白质上的过程是合子形成的重要环节,目前已证明附睾中分泌出来的一些特异蛋白质和精子成熟之间的相互关系,但精子动能的获得或与合子的结合能力的研究尚不十分清楚。     

南极鱼的“重生”——抗冻蛋白知多少

一条夏天出生的南极鱼第一次过冬,从妈妈那里初识它们过冬的法宝——抗冻蛋白,并了解了它们抗冻的原理。随后的抗冻比拼让它了解了多样的抗冻机制,以及抗冻蛋白在生活中广泛的应用。     

抗冻蛋白抗冻机制的分子模拟研究

抗冻蛋白能使生物体在寒冷环境下生存,具有极大的潜在应用价值。近年来,人们对抗冻蛋白开展了广泛的研究,但其抗冻机理还未明确。本文阐述了抗冻蛋白的功能特性和结构特征,并从结构的角度对其抗冻机制方面的分子模拟研究成果进行了综述。另一方面,对目前已知晶体结构的29个野生型抗冻蛋白的结构特性进行了分析,发现在整个抗冻蛋白表面和在冰结合位点处都存在亲水残基与水形成氢键和疏水残基与类冰结构特异性结合的特点。然后,探讨了抗冻蛋白的二级结构、冰结合位点残基的疏水性与抗冻活性之间的关系。最后,从结构的角度讨论了抗冻蛋白的机制和影响抗冻活性的因素并简要总结了仿生抗冻材料设计和应用的研究进展。     

短小芽孢杆菌碱性蛋白酶的结构研究

测定了短小芽孢杆菌碱性蛋白酶BP的分子量为19 500,分析了它的氨基酸组成。用N-溴代琥珀酰亚胺对酶进行了化学修饰,以紫外光谱法测得此酶含有2个色氨酸残基,其中一个残基为酶表现活性的必需基团,且位于分子表面。用荧光表面猝灭剂法推测另一个残基也位于分子表面或临近分子表面。通过其园二色谱证明此酶为无规则卷曲构象。     

操控冰水:仿生抗冻蛋白研究进展*

抗冻蛋白是自然界娴熟操控(冰)水的分子识别的典范之一。抗冻蛋白的抗冻活性与其特殊结构有着十分密切的关系。作为目前最高效的生物抗冻剂,抗冻蛋白因其含量低、易变性失活,导致产量过低,亟待开发新的来源。近年来,模拟抗冻蛋白的研究工作吸引了科学家们的广泛关注,抗冻蛋白关键的结构特质:氢键作用、疏水性、冰晶吸附、“结构水”在各类仿生抗结冰材料中相继得以体现,对深入理解抗冻蛋白作用机制起到了重要的推动作用。综述了仿生抗冻蛋白在仿生抗结冰材料领域的研究进展,对基于仿生抗冻蛋白的仿生抗结冰材料的发展做出了展望。     

抗冻蛋白与细胞色素蛋白P450水合层重定向动力学的比较分析

抗冻蛋白抑制冰结晶的机制仍不清晰. 例如,抗冻蛋白的水合水动力学及其与抗冻活性之间的关系尚无定论. 本文通过理论模拟,比较研究了来自云杉蚜虫的抗冻蛋白和细胞色素蛋白P450第一水合层中的水分子重定向动力学. 氢键供体水分子周围潜在受体水分子数量的增加导致水分子之间氢键交换的加速. 因此,抗冻蛋白抗冻活性区周围水分子的跳跃重定向动力学加速. 由于氢键交换的相互耦合和激发,随着氢键交换的加速,整个氢键网络的重排和水分子的框架重定向也被加速. 因此,抗冻蛋白的水分子重定向动力学比细胞色素蛋白P450更快. 本研究为理解抗冻蛋白提供了新的物理图像,并对抗冻蛋白的抗冻机理有了新的认识.     

IgG1型单克隆抗体对照品的一级结构解析

对单克隆抗体药物对照品(IgG1型)进行了全面的结构表征。采用ExactiveplusEMR质谱测定了去N糖前后抗体的精确分子量,并通过计算N糖含量得出其完整分子量为148285.0~149020.8,完整去糖后分子量为145810.4,N糖含量为1.67%~2.15%。优化了全柱成像实时等电聚焦毛细管电泳(WCID-cIEF)检测方法,结果显示该抗体等电点分布在8.21~8.74之间,其中主成分等电点为8.61,相对含量为61.43%。继而使用离子交换色谱检测了电荷异质性,发现碱性峰含量为4.1%,酸性峰含量为23.9%,主峰含量为72.0%,与WCID-cIEF结果吻合,并证明了方法间良好的重现性。通过切糖前后的肽图分析,获得糖肽分布信息,识别出糖修饰位点及重轻链的互补决定区（CDR）,甲硫氨酸(M)的氧化位点及氧化率,并通过抗体还原前后的肽图解析出二硫键连接。研究结果同时提示该抗体对照品无C末端糖化修饰现象。     

Purification and Primary Structure Determination of a Novel Polypeptide Isolated from Mistletoe Viscum coloratum

A novel polypeptide was isolated from mistletoe Viscum coloratum. The primarystructure of the polypeptide ‘named viscotoxin B2‘ was determined to be KSCCKNTTGRNIYNT CRFAGGSRERCAKLSGCKIISASTCPSDYPK by Edman degradation. Viscotoxin B2 shared highsequence homology with viscotoxins isolated from Viscum album. Pharmacological experimentsshowed that viscotoxin B2 had distinct cytotoxic activity on tumor cells. Viscotoxin B2 could beused as a leading compound in cancer therapy.     

Differential Scanning Calorimetric and Circular Dichroistic Studies on Plant Antifreeze Proteins

Antifreeze protein (AFP) can lower the freezing point by inhibiting the growth of ice crystals. In this article, the thermal hysteresis activity (THA) of a plant AFP was measured with differential scanning calorimetry (DSC). As is shown, when the amount of ice in the sample was less than 5% THA of this AFP reached as high as 0.35°C. The secondary structure of this AFP was studied with circular dichroism (CD). The CD spectrum from 195to 240 nm indicated a well-defined secondary structure consisting 11% α-helix, 34%antiparallel β-sheet and 55% random coil. This revised version was published online in August 2006 with corrections to the Cover Date.     

Interaction of Antifreeze Proteins with Hydrocarbon Hydrates

Recombinant antifreeze proteins (AFPs), representing a range of activities with respect to ice growth inhibition, were investigated for their abilities to control the crystal formation and growth of hydrocarbon hydrates. Three different AFPs were compared with two synthetic commercial inhibitors, poly‐N‐vinylpyrrolidone (PVP) and HIW85281, by using multiple approaches, which included gas uptake, differential scanning calorimetry (DSC) temperature ramping, and DSC isothermal observations. A new method to assess the induction period before heterogeneous nucleation and subsequent hydrate crystal growth was developed and involved the dispersal of water in the pore space of silica gel beads. Although hydrate nucleation is a complex phenomenon, we have shown that it can now be carefully quantified. The presence of AFPs delayed crystallization events and showed hydrate growth inhibition that was superior to that of one of the benchmark commercial inhibitors, PVP. Nucleation and growth inhibition were shown to be independent processes, which indicates a difference in the mechanisms required for these two inhibitory actions. In addition, there was no apparent correlation between the assayed activities of the three AFPs toward hexagonal ice and the cubic structure II (sII) hydrate, which suggests that there are distinctive differences in the protein interactions with the two crystal surfaces.     

Molecular basis for antifreeze activity difference of two insect antifreeze protein isoforms

The insect spruce budworm(Choristoneura fumiferana) produces antifreeze protein(AFP) to assist in the protection of the over-wintering larval stage and contains multiple isoforms. Structures for two isoforms,known as CfAFP-501 and CfAFP-337,show that both possess similar left-handed β-helical structure,although thermal hysteresis activity of the longer isoform CfAFP-501 is three times that of CfAFP-337. The markedly enhanced activity of CfAFP-501 is not proportional to,and cannot be simply accounted for,by the increased ice-binding site resulting from the two extra coils in CfAFP-501. In or-der to investigate the molecular basis for the activity difference and gain better understanding of AFPs in general,we have employed several different computational methods to systematically study the structural properties and ice interactions of the AFPs and their deletion models. In the context of intact AFPs,a majority of the coils in CfAFP-501 has better ice interaction and causes stronger ice lattice disruption than CfAFP-337,strongly suggesting a cooperative or synergistic effect among β-helical coils. The synergistic effect would play a critical role and make significant contributions to the anti-freeze activity β-helical antifreeze proteins. This is the first time that synergistic effect and its implica-tion for antifreeze activity are reported for β-helical antifreeze proteins.     

松香改性苯乙烯聚合物分子量及分子量分布的GPC测定

本文利用凝胶渗透色谱测定新型高分子材料（苯乙烯-歧化松香树脂乙烯酯）的分子量及分子量分布，实验数据重视性较好。标准偏差为0．074，变异系数为3．360％。     

Automated Protein Structure Elucidation From 2D NMR

A general method for automated de novo deduction of protein structure from 2D NMR has been developed. The algorithms which extract simple spin coupling topologies from MQF-COSY, construct more complicated spin coupling topologies based upon MQ spectrum including all possible pathways, and extract spin coupling topological fragments for amino acids of a protein, have been implemented in C~(++) language and run on a SUN 4/280 work station. Compared with the manual assignments for melittin, the total identity of the automated de novo method is 86.3%.     

Protein Secondary Structure Prediction with Partially Recurrent Neural Networks

Abstract

Partially recurrent neural networks with different topologies are applied for secondary structure prediction of proteins. The state of some activations in the network is available after a pattern presentation via feedback connections as additional input during the processing of the next pattern in a sequence. A reference data set containing 91 proteins in the training set and 15 non-homologous proteins in the test set is used for training and testing a network with a modified, hierarchical Elman architecture. The network predicts the secondary structures α-helix, β-sheet, and “coil” for each amino acid. The percentage of correctly classified amino acids is 67.83% on the training set and 63.98% on the test set. The best performance of a three-layer feedforward network is 62.7% on the same test set. A cascaded network, where the outputs of the recurrent network are processed by a second net with 13 × 3 inputs, four hidden and three output units has a predictive performance of 64.49%. The best corresponding feedforward net has a performance of 64.3%.     

Neural Networks Predict Protein Folding and Structure: Artificial Intelligence Faces Biomolecular Complexity

Abstract

In the genomic era DNA sequencing is increasing our knowledge of the molecular structure of genetic codes from bacteria to man at a hyperbolic rate. Billions of nucleotides and millions of aminoacids are already filling the electronic files of the data bases presently available, which contain a tremendous amount of information on the most biologically relevant macromolecules, such as DNA. RNA and proteins. The most urgent problem originates from the need to single out the relevant information amidst a wealth of general features. Intelligent tools are therefore needed to optimise the search. Data mining for sequence analysis in biotechnology has been substantially aided by the development of new powerful methods borrowed from the machine learning approach. In this paper we discuss the application of artificial feedforward neural networks to deal with some fundamental problems tied with the folding process and the structure-function relationship in proteins.     

Structure Analysis of Streptococcal Protein G Fc Binding Domain

The gene fragment (191 bp) encoding protein G IgG Fc binding domain was isolated by PCR from group G streptococcus (CMCC32138), and a clone containing this gene fragment was found to give fine reactivity to human IgG when expressed in Escherichia coli. The complete nucleotide sequence of the gene fragment was determined. One base pair differs from previously reported protein Gnucleotide sequences, and resultsin an amino acid change (Ala-Thr), but this variation makes no difference in binding to the IgG Fc part by ELISA.The secondary structure of the protein G IgG Fc binding domain has been estimated by circular dichroism and assigned by computer algorithm.It shows a typical α-helix region in this domain.By breaking this α-helix region with recombinant DNA techniques, a 44 peptide, which contained the N-terminal 27 amino acid residues of this domain, was expressed in E. coli and showed no reactivity to IgG.The hydropathicity of this domain was also analyzed and compared with that of protein A relevant