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1.
黑皮扁豆凝集素提纯及部分性质研究   总被引:2,自引:3,他引:2  
采用NaCl抽提、硫酸铵分部沉淀和2步离子交换法从黑皮扁豆中纯化得到一种甘露糖专一凝集素(DPL)。在非变性电泳中纯化的DPL只有一条蛋白带,在还原及非还原条件下的SDS-PAGE中呈现两条蛋白带,其分子量分别为36.8kD和39.8kD;利用FPLC经Sephacryl S-300凝胶过滤测得其表观分子量为155kD,表明DPL由2个α亚基和2个β亚基组成,无二硫键。DPL的等电点为pH6.18,含1.6%糖,为酸性糖蛋白。DPL的热稳定性和pH稳定性,与已报道的甘露糖/葡萄糖专一的豆科凝集素相似。  相似文献   

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以17β-雌二醇(E2)诱导雄性唐鱼产生卵黄蛋白原(Vtg),采用Sephacryl S-300凝胶过滤层析柱提纯了雄鱼整体匀桨液的Vtg。结果显示,已确定被纯化的唐鱼Vtg在4%—7.5% Native—PAGE电泳中分子量为440 kD左右,能同时被考马斯亮蓝、糖蛋白、磷蛋白、脂蛋白染色方法同时染色,是一种富含糖、磷、脂的蛋白。与对照组作比较,17β-雌二醇暴露组其体重明显下降(P<0.01),精巢发育滞后。结果表明,17β-雌二醇能够诱导雄性唐鱼产生大量的Vtg,并影响到其生长及精巢发育。雄性唐鱼Vtg可作为环境雌激素监测的有效生物学标记物。  相似文献   

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鬼臼多糖的分离纯化及其抗肿瘤活性   总被引:11,自引:1,他引:11  
鬼臼经乙醚脱脂后用热水抽提,用蛋白酶法和Sevag法相结合除去蛋白质,乙醇分级沉淀,经DEAE-Sepharose和Superdex G-75或Sephacryl S-200HR柱层析纯化得到两种多糖组分EPS-A和EPS-B。经分析该两种多糖均为单一组分,用分子筛层析法测定了EPS-A和EPS-B的分子量分别为15kD和175kD。纸层析和气相层析分析得知,EPS-B含有木糖,阿拉伯糖,鼠李糖,葡萄糖,甘露糖和半乳糖,其摩尔比为0.37:2.20:0.57:9.83:1.0:3.42,而EPS-A中仅含葡萄糖,体内试验结果表明,EPS-B多糖对小鼠腹水肝癌细胞生长有一定的抑制作用。  相似文献   

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介绍了从健康牛骨髓中分离到一种具有抑制肿瘤活性的糖蛋白,并对该糖蛋白进行了纯化,获得了电泳纯和层析纯的纯品.测得该糖蛋白N末端为两氨酸残基.用SDS-PAGE法测得该溏蛋白的表现分子量为65kD.氨基酸组成分析显示其为一种富含丝氨酸的蛋白质.生物活性测定表明,该糖蛋白纯品对小白鼠白血病P388细胞和人慢粒白血病HL-60细胞株的增殖均有抑制作用.有关该糖蛋白的糖含量的定量测定及蛋白质一级结构分析正在进一步的研究中.  相似文献   

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林生山黧豆谷氨酸脱羧酶的分离纯化及部分性质的研究   总被引:1,自引:0,他引:1  
以林生山黧豆为材料,利用硫酸按分段盐析,丙酮沉淀,DEAE-SepharoseFF离子交换柱层析,SephacrylS300凝胶过滤柱层析及FPLC-MonoQ柱层析技术,以聚酰胺薄膜层析荧光定量法为酶活力检测手段,分离纯化了谷氨酸脱羧酶,达到电泳银染纯.纯化后的林生山黧豆谷氨酸脱羧酶活力达375.09U·mp-1,纯化倍数38.2倍,经SDS-PAGE测定,其亚基分子量为70kD,经梯度PAGE确定,天然分子量为140kD,表明该酶是由两个亚基组成的二聚体.酶学研究表明,纯化的林生山黧豆谷氨酸脱羧酶的最适pH值为5.4,对谷氨酸的Km值为1.62×10-3mol·L-1,酶的最适温度为40℃,酶特异性地使谷氨酸脱羧,不能使天门冬氨酸等其它氨基酸脱羧.  相似文献   

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采用(NH4)2SO4沉淀,二次不同pH的DEAE-Sephacel离子交换柱层析及Sephacryl S-200分子筛柱层析从人癌性腹水中分离并纯化了PⅢP,并制备了抗血清。经SDS-PAGE鉴定为均一蛋白带,分子量约为47kD。经西德药盒测定,与西德、芬兰PⅢP抑制曲线平行关系良好。氨基酸组成测定亦与文献报道基本一致。抗血清效价较高,特异性强。本研究在方法学上有一定创新,提纯方法简便易行。  相似文献   

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采用丙酮粉抽提,DEAE-SephadexA-50、SephacrylS-300、MonoQ柱层析,从银杏(GinkgobilobaL.)花粉中分高纯化出微管蛋白(tubulin),其两个亚基(α、β)的分子量分别为54kD和52kD.纯化的微管蛋白可与鸡脑微管蛋白抗体发生免疫交叉反应.  相似文献   

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(一)本文用硫酸銨分段沉淀和DEAE纤維素柱层析方法从牛血浆中提取了一种对三氯乙酸不稳定的α_1-糖蛋白,經鉴定系超离心、电泳和免疫純。光散射法測定分子量为66,500,超离心-扩散法測定为67,000(偏微比容取0.65)。 (二)該蛋白质具有抑制胰蛋白酶的活力,酶与糖蛋白的克分子比值是2:1。 (三)該蛋白质与胰蛋白酶結合后,仍与其相应抗血清产生沉淀反应,文中对其抑制活力中心和抗原决定簇的关系作了討論。  相似文献   

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中药提取物中α-葡萄糖苷酶抑制剂的筛选   总被引:37,自引:1,他引:36  
目的:从中药中筛选α-葡萄糖苷酶抑制剂。方法:采用α-葡萄糖苷酶、淀粉酶以及蔗糖酶活性测定方法,对126种经水煮醇沉提取的常用中药进行α-葡萄糖苷酶抑制活性筛选。结果:在0.28mg/ml反应体系下,24种中药提取物显著抑制α-葡萄糖苷酶活性;在0.1mg/ml反应体系下,5种中药提取物抑制α-淀粉酶活性。大黄、山茱萸、赤芍、五倍子对α-葡萄糖苷酶、α-淀粉酶以及转化酶活性均有抑制作用,与阳性药物拜唐苹比较,这4种中药抑制α-淀粉酶的活性较低,但抑制α-葡萄糖计酶和转化酶的活性明显强于拜唐苹。结论:某些中药提取物能显著抑制α-葡萄糖苷酶活性,如果对其进一步分离、纯化并提取活性部位,可望获得抑制活性更强的中药α-葡萄糖苷酶抑制剂。  相似文献   

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《生命科学研究》2015,(5):392-396
用蒸馏水沉淀法、Sephacryl S-300 High Resolution Hiprep 26/60葡聚糖凝胶层析、蛋白质电泳的方法提取和纯化了拟环纹豹蛛Pardosa pseudoannulata的卵黄蛋白,并对其性质进行了分析。用蒸馏水沉淀法能得到卵黄蛋白粗提液,将得到的卵黄蛋白粗提液用Sephacryl S-300 High Resolution Hiprep 26/60凝胶柱过滤,可以得到纯化的卵黄蛋白。电泳结合不同的染色方法证明拟环纹豹蛛卵黄蛋白是一种糖脂复合蛋白,它由2个亚基组成,亚基的相对分子质量分别为80.1 k D和57.4 k D。  相似文献   

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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.  相似文献   

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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  相似文献   

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