中国鲎凝集素的分离纯化及性质研究

 经N-乙酰氨基葡萄糖交联琼脂糖亲和层析及以交联琼脂糖介质的高效液相分子筛层析,从中国鲎细胞溶解物中分离纯化了一种凝集素,其活性比原料鲎试剂提高128倍。鲎凝集素SDS电泳时表现出分子量为69000,和72000的二个亚基。N-乙酰氨基葡萄糖、D-半乳糖,D-甘露糖及岩藻糖等对鲎凝集素凝集鸡红细胞的活性有显著抑制作用,加热60℃,10分钟可使凝集素活性基本丧失。CaCl_2为凝集素活性所必需。鲎凝集素与肺炎球菌C多糖有沉淀反应。     

孔石莼(Ulya pertusa)凝集素的分离纯化及性质的研究

为抑制肿瘤细胞增殖和防治有关病害提供基础理论依据,将孔石莼(Ulva pertusa)经磷酸盐缓冲液抽提,20%～75%硫酸铵分级沉淀,牛甲状腺球蛋白-Sepharose 4B亲和层析,可以从绿藻孔石莼中纯化出孔石莼凝集素(UPL),在PAGE上显示单一蛋白染色带,在等电聚焦电泳上显示单一蛋白染色带,其pI为8.40.纯化后的UPL的最大紫外吸收峰在285 nm,用Sephadex G-200分子筛层析测得其分子量为11 047.该凝集素可以凝集人的A、B、AB、O型红细胞,且凝集活性相同,在对人(A、B、AB、O)兔、鲤、鲫的红细胞的凝集作用中,兔的凝集作用最强.该凝集素凝集兔红细胞的作用不被D-半乳糖、D-果糖、葡萄糖、蔗糖、甘露聚糖、γ球蛋白、卵清蛋白所抑制,仅被牛甲状腺球蛋白抑制,最小抑制浓度为6.20 g/L.该凝集素在pH4.0～10.14范围内均有活性,但在pH6.50～9.51范围内活性较高,该凝集活性在85℃加热1 h,活力仍未改变,说明具有很强的耐热性.     

重组人半乳凝集素-1的原核表达、纯化及生物活性检测

目的:在大肠杆菌中表达半乳凝集素-1(galectin-1),并进行纯化及生物活性检测。方法:将人半乳凝集素-1基因克隆至带有His融合标签的原核表达载体pQE-30上,转化大肠杆菌M15,经IPTG诱导表达,表达产物经亲和层析纯化后,进行Western印迹鉴定,并用红细胞凝集试验检测其生物学活性。结果:双酶切鉴定和核苷酸序列测定表明重组表达质粒pQE-30-Galectin-1构建正确;重组蛋白的表达量约占菌体总蛋白的50%,主要以可溶形式表达,纯化后蛋白纯度达95%以上,且具有良好的红细胞凝集活性。结论:在大肠杆菌中表达了重组人半乳凝集素-1,且具有良好的生物活性。     

甘薯凝集素的提取及性质研究

抗蔓割病甘薯的叶片组织浸取液,经硫酸铵分级沉淀,甲壳素柱层析及葡聚糖凝胶过滤得到一种在PAGE或SDS－PAGE上均呈现单一蛋白带的甘薯凝集素。该凝集素没有血型专一性及被测动物红细胞专一性,其凝集活性可被N－乙酰葡萄糖胺或岩藻糖所抑制。甘薯凝集素在75℃加热10min,即丧失全部凝集活性,其凝集活性依赖于Ca^2 和Mg^2 ,Mn^2 则无作用。经Sephadex G-100和SDS－PAGE测定,凝集素相对分子质量为63000,中性糖含量为6.21%,该凝集素对蔓割病菌有抑制作用,是一种酸性糖蛋白。     

蛋白核小球藻凝集素的分离纯化及部分性质研究

蛋白核小球藻藻粉的PBS抽提液经硫酸铵二步分级沉淀 ,再经DEAE Sepharose和SephadexG 10 0层析 ,从中分离纯化得到蛋白核小球藻凝集素 (CPL)。经测定 ,该凝集素为单个亚基的蛋白质 ,相对亚基分子量为 1 4× 10 4 — 1 5× 10 4 ,分子中不含糖。在氨基酸组成中 ,苯丙氨酸 (Phe)的含量最高 ,其次是天冬氨酸 (Asp)和谷氨酸 (Glu) ,不含组氨酸 (His)。CPL能够凝集兔、绵羊及鸽子红细胞 ,其中对兔红细胞的凝集活性最大 ,最低浓度为 6 88μg/mL ,对鸡、鸭及人红细胞 (A型、O型及B型 )无凝集活性。卵黏蛋白和 7种单糖对CPL的凝血活性具有抑制作用。CPL具有很好的热稳定性 ,在 90℃处理 10min不失活。     

孔石莼(Ulva pertusa)凝集素的分离纯化及性质的研究

为抑制肿瘤细胞增殖和防治有关病害提供基础理论依据 ,将孔石莼 (Ulva pertusa)经磷酸盐缓冲液抽提 ,2 0 %～ 75%硫酸铵分级沉淀 ,牛甲状腺球蛋白 - Sepharose4B亲和层析 ,可以从绿藻孔石莼中纯化出孔石莼凝集素 (UPL) ,在 PAGE上显示单一蛋白染色带 ,在等电聚焦电泳上显示单一蛋白染色带 ,其 p I为 8.40 .纯化后的 UPL的最大紫外吸收峰在 2 85nm,用 Sephadex G- 2 0 0分子筛层析测得其分子量为 1 1 0 4 7.该凝集素可以凝集人的 A、B、AB、O型红细胞 ,且凝集活性相同 ,在对人 (A、B、AB、O)兔、鲤、鲫的红细胞的凝集作用中 ,兔的凝集作用最强 .该凝集素凝集兔红细胞的作用不被 D-半乳糖、D-果糖、葡萄糖、蔗糖、甘露聚糖、γ球蛋白、卵清蛋白所抑制 ,仅被牛甲状腺球蛋白抑制 ,最小抑制浓度为 6.2 0 g/L.该凝集素在 p H4.0～ 1 0 .1 4范围内均有活性 ,但在p H6.50～ 9.51范围内活性较高 ,该凝集活性在 85℃加热 1 h,活力仍未改变 ,说明具有很强的耐热性 .     

江苏菜豆同工凝集素的分离纯化及性质研究

江苏菜豆经酸水(PH2.0)抽提,硫酸铵分级沉淀,分离植物血球凝集素(PHA-P),分子量为128000的糖蛋白,活性回收率在80％以上,PHA-P经SP-sephadexc-50离子交换层析,分成L_4,L_3E_1,L_2E_2,L_1E_3,和E_4同工凝集素。 L_4和E_4等电点为5.4和6.5。亚基分子量分别是31000和33000,并有类似的氨基酸组成。PAGE分析为单一蛋白带。红细胞凝集活性随电泳迁移速度的加快而增强,促淋性细胞分裂活性则减弱。E_4血凝活性受CalNAc,EDTA抑制和Zn~(++)的促进。     

一种新的蘑菇凝集素的生化特征和氨基酸分析

用 (NH4) 2 SO4分级沉淀、离子交换和分子筛等方法 ,从食用蘑菇中分离纯化出一种凝集素 ,经SDS -PAGE测定其亚基的相对分子量为 15 .8kDa,质谱分析法测定其分子量为 32kDa ,说明该凝集素由两个亚基组成。氨基酸分析发现 ,该凝集素不含半胱氨酸。热稳定试验和耐酸碱试验显示 ,该凝集素活性具很高稳定性     

蛇毒C-型凝集素研究进展

蛇毒中含有丰富的非酶活性C－型凝集素蛋白,根据其结构及功能的差异,该类蛋白可分为Ca^2 依赖的有糖基识别活性的C－型真凝集素及无糖基识别活性的C－型凝集素样蛋白。C－型真凝集素的结构相似度高,而功能却较为单一,具有特异性糖结合活性;C－型凝集样蛋白的结构变异度大,活性亦具有多样性。后者能通过特异性的蛋白质－蛋白质相互作用而作用于血液凝固系统及血小板,从而发挥抗凝或促凝的生理功能。     

狗脊蕨凝集素的分离纯化与部分性质

狗脊蕨叶组织经磷酸缓冲液抽提,硫酸铵分级沉淀,DEAE-Sepharose(fast flow)离子交换层析和Sephadex G-100分子筛层析,获得具有凝集活性的狗脊蕨凝集素.用PAGE和SDS-PAGE检测均显示1条蛋白带,提示狗脊蕨凝集素分子只有一个亚基.Sephadex G-100层析和SDS-PAGE测得其相对分子质量在20 kD左右.该凝集素对红细胞有种属专一性,其凝集活性可被木糖、卵粘蛋白所抑制.狗脊蕨凝集素对热相当稳定,以100℃温度加热10 min仍保存部分凝集活性.凝集活性不依赖于金属离子Mg2 、Ca2 、Mn2 .它的中性糖含量为3.1%,Phe含量较高,His含量较低,不含Tyr和Pro.     

Investigation of the mechanism of temperature sensitivity of the electroreceptors of ampullae of lorenzini

In experiments on Black Sea skates (Raja clavata), the potential of the receptor epithelium of the ampullae of Lorenzini and spike activity of single nerve fibers connected to them were investigated during electrical and temperature stimulation. Usually the potential within the canal was between 0 and –2 mV, and the input resistance of the ampulla 250–400 k. Heating of the region of the receptor epithelium was accompanied by a negative wave of potential, an increase in input resistance, and inhibition of spike activity. With worsening of the animal's condition the transepithelial potential became positive (up to +10 mV) but the input resistance of the ampulla during stimulation with a positive current was nonlinear in some cases: a regenerative spike of positive polarity appeared in the channel. During heating, the spike response was sometimes reversed in sign. It is suggested that fluctuations of the transepithelial potential and spike responses to temperature stimulation reflect changes in the potential difference on the basal membrane of the receptor cells, which is described by a relationship of the Nernst's or Goldman's equation type.I. P. Pavlov Institute of Physiology, Academy of Sciences of the USSR, Leningrad. I. M. Sechenov, Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Pacific Institute of Oceanology, Far Eastern Scientific Center, Academy of Sciences of the USSR, Vladivostok. Translated from Neirofiziologiya, Vol. 12, No. 1, pp. 67–74, January–February, 1980.     

Principles of organization and evolution of systems of regulation of functions

Evolution of living organisms is closely connected with evolution of structure of the system of regulations and its mechanisms. The functional ground of regulations is chemical signalization. As early as in unicellular organisms there is a set of signal mechanisms providing their life activity and orientation in space and time. Subsequent evolution of ways of chemical signalization followed the way of development of delivery pathways of chemical signal and development of mechanisms of its regulation. The mechanism of chemical regulation of the signal interaction is discussed by the example of the specialized system of transduction of signal from neuron to neuron, of effect of hormone on the epithelial cell and modulation of this effect. These mechanisms are considered as the most important ways of the fine and precise adaptation of chemical signalization underlying functioning of physiological systems and organs of the living organism