Pharmacologically active spider peptide toxins

Advances in mass spectrometry and peptide biochemistry coupled to modern methods in electrophysiology have permitted the isolation and identification of numerous novel peptide toxins from animal venoms in recent years. These advances have also opened up the field of spider venom research, previously unexplored due to methodological limitations. Many peptide toxins from spider venoms share structural features, amino acid composition and consensus sequences that allow them to interact with related classes of cellular receptors. They have become increasingly useful agents for the study of voltage-sensitive and ligand-gated ion channels and the discrimination of their cellular subtypes. Spider peptide toxins have also been recognized as useful agents for their antimicrobial properties and the study of pore formation in cell membranes. Spider peptide toxins with nanomolar affinities for their receptors are thus promising pharmacological tools for understanding the physiological role of ion channels and as leads for the development of novel therapeutic agents and strategies for ion channel-related diseases. Their high insecticidal potency can also make them useful probes for the discovery of novel insecticide targets in the insect nervous system or for the development of genetically engineered microbial pesticides.Received 19 March 2003; received after revision 9 May 2003; accepted 16 May 2003     

Deamidation and isoaspartate formation in proteins: unwanted alterations or surreptitious signals?

Formation of -linked Asp-Xaa peptide bonds – isoaspartyl (isoAsp) sites – arise in proteins via succinimide–linked deamidation of asparagine or dehydration of aspartate, reactions which represent a major source of spontaneous protein damage under physiological conditions. Accumulation of atypical isoaspartyl sites is minimized in vivo by the activity of protein L-isoaspartyl O–methyltransferase (PIMT), which regenerates a normal peptide bond. Loss of PIMT has harmful consequences, especially in neurons; thus, formation of isoAsp sites and their subsequent correction by PIMT is widely believed to constitute an important pathway of protein damage and repair. Recent evidence is mounting, however, that deamidation and isoaspartate formation may, in some instances, constitute a novel mechanism for intentional modification of protein structure. Herein we describe the mechanism of Asx rearrangement, summarize the evidence that PIMT serves an important repair function, and then focus on emerging evidence that deamidation and isoAsp formation may sometimes have a useful function.Received 16 October 2002; received after revision 11 December 2002; accepted 12 December 2002     

In pre-sterol carrier protein 2 (SCP2) in solution the leader peptide 1 – 20 is flexibly disordered, and residues 21 – 143 adopt the same globular fold as in mature SCP2

The preform of the rabbit sterol carrier protein 2 (pre-rSCP2) was cloned, the uniformly ¹⁵N-labelled protein expressed in Escherichia coli and studied by three-dimensional ¹⁵N-resolved nuclear magnetic resonance spectroscopy. In spite of its low solubility in aqueous solution of only ∼0.3 mM, sequential ¹⁵N and ¹H backbone resonance assignments were obtained for 105 out of the 143 residues. From comparison of the sequential and medium-range nuclear Overhauser effects (NOEs) in the two proteins, all regular secondary structures previously determined in mature human SCP2 (hSCP2) [Szyperski et al. (1993) FEBS Lett. 335: 18–26] were also identified in pre-rSCP2. Near-identity of the backbone ¹⁵N and ¹H chemical shifts and 1 : 1 correspondence of 24 long-range NOEs to backbone amide groups in the two proteins show that the residues 21 – 143 adopt the same globular fold in pre-rSCP2 and mature hSCP2. The N-terminal 20-residue leader peptide of pre-rSCP2 is flexibly disordered in solution and does not observably affect the conformation of the polypeptide segment 21 – 143. Received 11 May 1998; accepted 15 May 1998     

噬菌体肽库技术及应用

噬菌体肽库技术是近年来发展起来的一项重要技术,具有广泛的应用前景.本文综述了这一技术的发展以及它在各个领域内的应用.     

从吉林林蛙干皮中提取抗菌肽

从吉林林蛙干皮中分离提取了具有广谱抗菌作用的多肽 ,制备得到抗菌肽样品。在实验中摸索出提取抗菌肽的最佳工艺路线 ,建立了抗菌肽的纯化方法 ,并对所得到的抗菌肽的氨基酸进行组成分析     

海洋活性物质C03抑菌及抗流感病毒的作用

活性物质CO3是从海洋分离出一种相对分子质量小于5000的抑菌肽，该物质对金黄色葡萄球菌(26001)等多种传染细菌及流感病毒鼠肺适应株FM1，表现出具有较好的抑菌及抗病毒作用，可望开发一种抗菌及抗病毒海洋新药。     

杆状病毒表达的hZP3 B细胞表位嵌合肽的免疫活性研究

为研究杆状病毒系统表达的人透明带ZP3的B细胞表位嵌合肽(BCP)的免疫原性及其抗血清与人透明带的免疫交叉反应，重组z3bcp病毒用于感染sf9细胞以表达BCP。Tricine—SDS—PAGE用于分离感染液中的小相对分子质量表达产物，表达产物用于免疫小鼠制备抗血清，免疫细胞化学试验用于检验抗血清与人卵泡透明带的交叉反应性。结果显示，矽细胞被z3bcp病毒感染后，培养液中分离出一条相对分子质量约为6000的特异带，而正常矽细胞中没有等同的带。免疫细胞化学试验结果显示，该特异肽的抗血清能与人透明带特异结合，表明人透明带BCP嵌合肽在杆状病毒系统中成功表达，表达产物具有抗原性，抗血清能与人透明带特异结合。     

禾谷炭疽菌中候选G蛋白偶联受体蛋白理化性质及遗传关系分析

GPCR是G蛋白偶联受体的简称,作为一类具有七跨膜螺旋结构的重要细胞表面受体,对生物中细胞信号转导过程发挥着重要作用。然而,对于禾谷炭疽菌候选GPCR蛋白的理化性质及遗传关系尚不清楚,制约着对该菌致病机制的研究。本研究基于对禾谷炭疽菌中220个候选GPCR蛋白序列,利用SignalP 5.1、ProtParam和ProtScal等在线预测程序对上述蛋白中的信号肽、理化性质、疏水性进行明确,结果表明,禾谷炭疽菌中仅有7个含有信号肽的候选GPCR蛋白,理论等电点主要集中在大于6.01,多属于中性或碱性蛋白,均为疏水性蛋白,同时利用MEGA软件对上述蛋白的遗传关系进行解析,明确候选GPCR蛋白主要分为A、B、C三类,上述类别与其保守结构域有着密切关系。该研究为进一步开展禾谷炭疽菌GPCR蛋白功能解析提供了重要的理论支撑,也为其他炭疽菌中GPCR蛋白功能研究提供了重要的理论基础。     

神经元及神经环路活性异常与阿尔茨海默病研究进展

 细胞外淀粉样蛋白（Aβ）沉积和细胞内神经纤维缠结是阿尔茨海默病的典型病理特征。淀粉样蛋白和tau蛋白（神经纤维缠结的主要组成成分）在脑中的异常聚集会导致神经元活性异常,进而引起神经环路结构及功能紊乱,最终造成阿尔茨海默病患者认知功能障碍。概述了Aβ及tau蛋白的生成及调控,阐述了Aβ及tau蛋白异常聚集在神经元及神经环路活动中的作用和机制,综述了ApoE、炎症反应及成体神经发生异常在AD神经元及神经环路活动障碍中的作用。