R9-FOXM1(1-234aa)重组蛋白批量纯化及其抑瘤效应研究

通过基因工程构建重组表达质粒pET15b-R9-FOXM1(1-234aa),转化大肠杆菌建立构建表达R9-FOXM1(1-234aa)的菌株.采用原核表达系统和His-tag亲和纯化手段,规模化制备纯化穿膜肽R9-FOXM1(1-234aa),获得的蛋白的纯度达到90%以上.用R9-FOXM1(1-234aa)穿膜肽处理不同的肿瘤细胞,通过MTT实验研究其细胞效应.结果显示:当R9-FOXM1(1-234aa)穿膜肽的浓度达到2mM时,肿瘤细胞的死亡率为50%左右.实验表明穿膜肽R9-FOXM1(1-234aa)抑制不同肿瘤细胞的生长,有可能成为治疗肿瘤的潜在蛋白类药物.     

基于免疫学的入侵检测系统模型

随着网络安全问题日益突出 ,入侵检测越来越受到关注 ,针对目前各种类型的防火墙和防病毒软件都存在一定的缺陷 ,该文基于仿生学的免疫原理 ,将肽链定义为在网络操作系统中由授权程序执行的系统调用短序列 ,提出了一种新型的入侵检测系统———基于免疫学的入侵检测系统 ,并对其主要功能模块 :免疫计算机和监控器进行了分析。该系统可有效地提高系统实时检测和响应入侵的能力     

4种促性腺激素释放激素抗体的制备、鉴定和纯化

用3-马来酰亚胺基苯甲酸琥珀酰亚胺酯(MBS)法将人工合成的4种不同来源的促性腺激素释放激(GnRH)(即章鱼GnRH、海鞘GnRH-Ⅰ、七鳃鳗GnRH-Ⅰ和七鳃鳗GnRH-Ⅲ)分别与匙孔戚血蓝蛋白(KLH)偶联后作为抗原,免疫新西兰大白兔获得GnRH抗体.用间接ELISA方法检测抗血清效价,并用Western blot鉴定其特异性.制备抗原肽亲和纯化柱,纯化多克隆抗体.结果显示,4种多克隆抗体效价高达1∶500 000(体积比),并能和相应的GnRH多肽特异性结合,纯化后的抗体在SDS-PAGE显示为单一条带.通过上述方法,获得4种高效价、高纯度的GnRH抗体,为低等动物GnRH的研究提供了理论依据.     

文蛤蛋白水解制备抗氧化活性肽的研究

以文蛤蛋白为原料,通过蛋白酶水解制备抗氧化活性肽.利用体外清除DPPH活性评价酶解产物的抗氧化活性,采用超滤、葡聚糖凝胶柱层析和HPLC对相应的活性肽进行分离纯化,通过HPLC-MS进行结构鉴定.结果表明:木瓜蛋白酶水解产物的DPPH清除活性明显高于碱性蛋白酶、风味蛋白酶、中性蛋白酶和复合蛋白酶,产物质量浓度为1 mg·mL-1时,DPPH清除率为55.74%;以DPPH清除率为筛选指标,对木瓜蛋白酶酶解产物进行逐级分离,得到4个具有抗氧化活性的目的肽段,氨基酸序列分别为LNFNLEKSR(1120.3 u)、SWLRPR(814.4 u)、RIGNIISQY(1063.3 u)和LNFNLEK(877.2 u).     

Effects of enterally- and parenterally-administered bombesin on intestinal luminal tryptic activity and protein in the suckling rat

Summary Because of the presence of bombesin-like immunoreactivity in milk we investigated if enteral administration of bombesin affects the intestinal luminal content of trypsin and protein in 12-14-day-old rats. Bombesin (40 g/kg), given either orogastrically or subcutaneously, produced a significant elevation in the intestinal content of trypsin activity. Thus, enterally-administered bombesin can produce acute biologic effects in suckling rats.     

Synthesis ofO 1.5-(β-D-galactopyranosyl) [DMet2, Hyp5] enkephalin amide,a new highly potent analgesic enkephalin-related glycosyl peptide

Summary A new series of O-glycosyl enkephalins has been prepared, following a convergent strategy, with high chemical yields. The galactosyl analogue,O ^1.5-(-D-galactopyranosyl) [DMet², Hyp⁵] enkephalin amide proved to be one of the most potent in vivo opioid agonists synthesized up to now.     

Control of the cardiovascular system ofAplysia by identified neurons

The neural network that controls the cardiovascular system ofAplysia adapts cardiovascular function to a variety of different physiological and behavioral situations. It (1) coordinates the cardiovascular system with the renal and respiratory systems; (2) modifies both systemic and regional blood flow during food-elicited arousal and feeding; and (3) changes the tension of longitudinal vascular muscle to adapt the arterial tree to changes in body shape. Indirect evidence suggests that the cardiovascular control circuit may also play a role in maintaining homeostasis during egg laying. Several putative neurotransmitters, including acetylcholine, serotonin, R151 and R152 peptides, have been localized to identified neurons in this circuit.     

Regulatory peptide immunocytochemistry at light- and electron microscopical levels

Summary Immunocytochemical techniques applied at both light- and electron microscopical levels are valuable in the study of regulatory peptide distribution in normal and diseased tissue, whether in the form of sections or whole cell preparations. Successful immunolocalisation depends on 1) adequate preservation of the peptide antigen and the tissue structure in which it resides; 2) a suitably specific and sensitive labelled antibody detecting system. In general, peptides are stable molecules, most of which retain their antigenicity after conventional cross-linking fixation and tissue processing, allowing standard immunocytochemical methods to be used for light- and electron microscopy. Regulatory peptides are derived from precursor molecules and several families of structurally similar peptides are now generally recognised. Region-specific antibodies may be needed to overcome problems of cross-reactivity or to identify a bioactive form in the presence of its precursor. Multiple co-localisation of different related and unrelated peptides in the same cell or even storage granule is now recognised and can be identified by immunocytochemistry.     

Cephalopod myocardial receptors: Pharmacological studies on the isolated heart ofSepia officinalis (L.)

Summary This paper presents a synopsis of the information available about the pharmacological action of various substances on the cephalopod heart, with special emphasis on the central heart ofSepia officinalis. Threshold concentrations, EC₅₀ values and maximum effective concentrations have been experimentally determined. Studies with various transmitter substances, analogous compounds and antagonists have led to the following picture: Acetylcholine is the natural inhibitory transmitter substance; it acts via receptors with nicotinic properties which can be blocked by d-tubocurarine and -bungarotoxin. The probable excitatory transmitter system is represented by a noradrenergic innervation. Noradrenaline has a positive inotropic and a positive chronotropic action on in vitro heart preparations. A positive inotropic response can also be evoked by serotonin (5-HT); this effect is not due to stimulation of the catecholamine receptor, as is shown by cross-over experiments with specific blocking agents. Furthermore, a peptidergic receptor system has been described which reacts with the molluscan cardioactive peptide FMRF amide most effectively. It is assumed that cardioactive peptides may reach the central heart in the circulating blood; the sites of synthesis and release are still unknown. Possibly the NSV-layer of the vena cava is involved in hormonal cardiovascular regulation processes.     

小分子蛋白酶抑制剂及多肽毒素的研究回顾

对以往小分子蛋白酶抑制剂及多肽毒素的研究成果作一历史性回顾, 重点介绍了Bowman-Birk家族的绿豆胰蛋白酶抑制剂, 对此抑制剂的Lys活性功能区作了解析. 它是双头抑制剂, 有两个功能区, 用蛋白与基因测序、基因表达、突变、肽段合成等手段证实, 其核心结构是一个由两个相连九元环组成的16肽, 与天然14环肽的向日葵抑制剂极类似. 另一深入研究的抑制剂是27肽的天花粉胰蛋白酶抑制剂, 属于南瓜族, 介绍了该家族的基因序列. 多肽毒素的研究包括蝎毒毒素及芋螺毒素. 本文介绍了多种东亚马氏钳蝎新的钾通道毒素, 有的是钙离子依赖的BK或SK钾通道毒素, 有的是电压门控毒素, 有的具有两个不同的功能区, 阐明了它们的构效关系. 表达的重组蝎氯毒素有望用于恶性胶质瘤的治疗. 芋螺毒素分子量小、序列多变、功能广泛, 有更好的应用前景. 从中国南海16种芋螺中纯化了50个结构新的芋螺毒素, 克隆了134个新基因; 确定了3个新超家族, 命名为V, Y, K超家族; 研究发现, 在某些芋螺毒素一级结构中有新的二硫键骨架和二硫键配对、独特的模板(motif)、氨基酸的D型化等; 分析了A超家族的基因组结构, 在内含子的3′端有一非常保守的序列, 可用作引物克隆更多新的芋螺毒素基因; 阐明了个别芋螺毒素的生理功能.