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Isolation of populations of antipeptide antibodies directed against different epitopes of the same fragment. 针对同一片段不同表位的抗肽抗体群体的分离。
Pub Date : 1985-01-01 DOI: 10.1515/znc-1985-1-230
A Chersi, C Greger, R A Houghten

Rabbit antibodies against small peptides may be composed by subpopulations recognizing different epitopes made likely by few amino acids. This explains the frequent crossreactivity of antipeptide antibodies with unrelated peptides. A suitable use of immunoadsorbents is suggested to obtain truly specific antibodies able to react with restricted amino acid sequences.

兔抗小肽抗体可能由识别不同表位的亚群组成,这些表位可能是由很少的氨基酸组成的。这解释了抗肽抗体与不相关肽的频繁交叉反应性。建议适当使用免疫吸附剂以获得能够与限制性氨基酸序列反应的真正特异性抗体。
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引用次数: 0
In vitro cellular muscle calcium metabolism. Characterization of effects of 1,25-dihydroxy-vitamin D3 and 25-hydroxy-vitamin D3. 体外细胞肌钙代谢。1,25-二羟基维生素D3和25-羟基维生素D3作用的表征。
Pub Date : 1985-01-01 DOI: 10.1515/znc-1985-1-220
A R de Boland, R Boland

Cultures of vitamin D-deficient chick soleus muscle and 12 day-old chick embryo myoblasts were used to characterize the effects of 1,25-dihydroxy-vitamin D3 and 25-hydroxy-vitamin D3 on muscle cell Ca metabolism. Physiological amounts of both sterols increased the rate and extent of 45Ca uptake by cultures. However, 1,25(OH)2D3 was significantly more effective than 25 OHD3. The greater potency of 1,25(OH)2D3 to increase Ca uptake could be shown after various treatment intervals of cultures and using a wide concentration range of both derivatives. Information about Ca pools affected by vitamin D3 metabolites was obtained through kinetic analysis of Ca efflux in cultured myoblasts. Cytoplasmic and mitochondria Ca pools were identified on the basis of their half-times of desaturation and by selective inhibition of plasma membrane and mitochondrial Ca transport with LaCl3 and Ruthenium Red, respectively. The data suggests that 1,25(OH)2D3 acts on muscle cellular Ca by increasing Ca efflux and influx through mitochondrial and plasma membranes whereas the predominant effect of 25 OHD3 is to increase Ca influx into mitochondria.

采用维生素d缺乏鸡比目鱼肌和12日龄鸡胚成肌细胞培养,研究1,25-二羟基维生素D3和25-羟基维生素D3对肌肉细胞钙代谢的影响。两种甾醇的生理量增加了培养物吸收45Ca的速度和程度。然而,1,25(OH)2D3明显比25 OHD3更有效。125 (OH)2D3在不同的培养处理间隔和使用两种衍生物的宽浓度范围后,可以显示出更大的增加钙吸收的效力。通过对培养成肌细胞钙外排的动力学分析,获得了维生素D3代谢物对钙池影响的信息。细胞质和线粒体的钙池分别根据它们的半饱和时间和LaCl3和钌红选择性抑制质膜和线粒体钙运输来确定。数据表明,1,25(OH)2D3通过增加钙通过线粒体和质膜的外排和内流来作用于肌肉细胞钙,而25 OHD3的主要作用是增加钙流入线粒体。
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引用次数: 19
Enzymatic nitrate assay by a kinetic method employing Escherichia coli nitrate reductase. 用大肠杆菌硝酸还原酶动力学法测定硝酸盐。
Pub Date : 1985-01-01 DOI: 10.1515/znc-1985-1-226
J Schild, J H Klemme

An enzymatic assay system for nitrate employing the membrane-bound nitrate reductase (EC 1.7.99.4) of E. coli is described. Contrary to previous enzymatic assay systems, the present method is a kinetic one, i.e. the substrate, nitrate, is assayed by measuring the reaction rate of the nitrate reductase-catalyzed reaction. Based on the observation that the nitrate reductase-catalyzed reaction obeys pseudo-first order kinetics, a test system is described allowing the assay of nitrate at a concentration as low as 1 ppm. The relatively high Michaelis-Menten constant for nitrate (0.3 mM) of the E. coli nitrate reductase favours nitrate assay by the kinetic method.

描述了一种利用大肠杆菌的膜结合硝酸盐还原酶(EC 1.7.99.4)的硝酸盐酶测定系统。与以前的酶分析系统相反,本方法是一种动力学方法,即通过测量硝酸还原酶催化反应的反应速率来测定底物硝酸盐。基于对硝酸还原酶催化反应服从准一级动力学的观察,描述了一种测试系统,允许在低至1ppm的浓度下测定硝酸盐。大肠杆菌硝酸盐还原酶的Michaelis-Menten常数较高(0.3 mM),有利于用动力学法测定硝酸盐。
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引用次数: 5
Chemical and physical characterization of four interfacial-active rhamnolipids from Pseudomonas spec. DSM 2874 grown on n-alkanes. 在正构烷烃上生长的伪单胞菌DSM 2874中四种界面活性鼠李糖脂的化学和物理性质。
Pub Date : 1985-01-01 DOI: 10.1515/znc-1985-1-212
C Syldatk, S Lang, F Wagner, V Wray, L Witte

Four extracellular glycolipids produced under growth-limiting conditions were isolated from the culture broth of Pseudomonas spec. DSM 2874. After purification by column and thick-layer chromatography they were identified as anionic rhamnolipids. 1H and 13C-NMR studies showed that two of these, beta(beta(2-O-alpha-L-rhamnopyranosyloxy)decanoyl)decanoic acid and beta(beta(2-O-alpha-L-rhamnopyranosyl-alpha-L-rhamnopyranosylox y)decanoyloxy) decanoic acid, were identical with compounds described previously, while the other more hydrophilic compounds, beta(2-O-alpha-L-rhamnopyranosyloxy)decanoic acid and beta(2-O-alpha-L-rhamnopyranosyl-alpha-L-rhamnopyranosyloxy) decanoic acid, were new compounds. Surface and interfacial activity of the organic crude extract and of the purified components were determined in different aqueous solutions. The pH-dependence of surface and interfacial properties of the two previously described rhamnolipids (4, 20, 23) were examined in Teorell-Stenhagen-buffer (supplemented with 10% NaCl) at pH 3.0 and pH 9.0. All rhamnolipids reduced the surface-tension from 72 to about 30 mN/m and the interfacial-tension from 42 to about 1 mN/m. The critical micelle concentrations were of the order of 5 to 200 mg/l depending on the structure of the molecule.

从假单胞菌DSM 2874培养液中分离出生长受限条件下产生的4种胞外糖脂。经柱层析和厚层色谱纯化,鉴定为阴离子鼠李糖脂。1H和13C-NMR研究表明,其中β (β (2- o - α - l- rhamnopyranosyloxy)癸酸和β (β (2- o - α - l- rhamnopyranosyloxy)癸酸与先前描述的化合物相同,而其他更亲水的化合物β (2- o - α - l- rhamnopyranosyloxy)癸酸和β (2- o - α - l- rhamnopyranosyloxy)癸酸是新化合物。测定了有机粗提物和纯化组分在不同水溶液中的表面和界面活性。在pH 3.0和pH 9.0的teorel - stenhagen缓冲液(添加10% NaCl)中,研究了上述两种鼠李糖脂(4,20,23)的表面和界面性质对pH的依赖关系。所有鼠李糖脂都将表面张力从72降低到约30 mN/m,界面张力从42降低到约1 mN/m。根据分子结构的不同,胶束的临界浓度为5 ~ 200mg /l。
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引用次数: 181
Constituents of Agaricus xanthodermus Genevier: the first naturally endogenous azo compound and toxic phenolic metabolites. 黄皮蘑菇的成分:第一种天然内源性偶氮化合物和有毒酚类代谢物。
Pub Date : 1984-11-01 DOI: 10.1515/znc-1984-11-1203
M Gill, R J Strauch

Extraction of fresh sporophores of the fungus Agaricus xanthodermus yields 4,4'-dihydroxy-azobenzene, phenol, p-quinol, and 4,4'-dihydroxybiphenyl. This is the first report of an azo compound arising endogenously in nature, while phenol, p-quinol and 4,4'-dihydroxybiphenyl have not previously been isolated from higher fungi. Phenol is present in fruitbodies in sufficiently high concentration to account for the toxicity of A. xanthodermus.

从真菌黄皮蘑菇的新鲜孢子中提取出4,4′-二羟基偶氮苯、苯酚、对喹啉和4,4′-二羟基联苯。这是首次从自然界中分离到偶氮化合物,而苯酚、对喹啉和4,4′-二羟基联苯此前尚未从高等真菌中分离到。酚存在于子实体中,其浓度足够高,足以说明黄皮霉的毒性。
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引用次数: 36
Dependence on membrane lipids of the effect of vanadate on calcium and ATP binding to sarcoplasmic reticulum ATPase. 钒酸盐对钙和ATP与肌浆网ATP酶结合作用的膜脂依赖性。
Pub Date : 1984-11-01 DOI: 10.1515/znc-1984-11-1224
P Medda, W Hasselbach

The affinity of the sarcoplasmic reticulum transport ATPase for calcium and ATP is not affected by lipid deprivation while vanadate binding is completely abolished. Lipid substitution restores vanadate binding as well as the vanadate induced disappearance of the enzyme's high affinity calcium and nucleotide binding sites. Nucleotide binding is simultaneously restored with the displacement of vanadate from the enzyme following the occupation of its low affinity calcium binding sites.

肌浆网转运ATP酶对钙和ATP的亲和力不受脂质剥夺的影响,而钒酸盐的结合则完全被取消。脂质取代恢复了钒酸盐结合以及钒酸盐诱导的酶的高亲和力钙和核苷酸结合位点的消失。核苷酸结合同时恢复,钒酸盐从占据其低亲和力钙结合位点的酶的位移。
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引用次数: 3
Preparation of an affinity chromatographic system for the separation of ADP binding proteins. ADP结合蛋白亲和色谱分离系统的制备。
Pub Date : 1984-11-01 DOI: 10.1515/znc-1984-11-1207
E Bieber, C Woenckhaus, H Pauli

[4-(3-Bromoacetylpyridinio)-butyl]adenosine pyrophosphate as a structural analog of NAD+ reacts covalently with the sulfhydryl groups of thiopropyl agarose. 10-20 mumol can be bound to 1 ml gel. Stabilization of the insoluble coenzyme is attained by treatment with sodium boro hydride (NaBH4). This complex when applied to column chromatography, allows the separation of various dehydrogenases as a result of their different complex stability coefficients. Alcohol dehydrogenase from liver, lactate dehydrogenase, and adenylate kinase, which all bind to the ADP-analog residues of the gel matrix, can thus be separated by different salt gradients. Alcohol dehydrogenase from yeast, however, does not form a complex and can easily be eluted from the column with phosphate buffer. Glyceraldehyde-3 phosphate and aldehyde dehydrogenases can be eluted by the addition of NAD+ or NADH to the buffer. The uncharged 1,4-dihydropyridine ring of the reduced coenzyme produces a more stable complex with the dehydrogenases than the oxidized form.

[4-(3-溴乙酰吡啶)-丁基]焦磷酸腺苷作为NAD+的结构类似物与硫丙基琼脂糖的巯基共价反应。10-20 μ mol可与1ml凝胶结合。不溶性辅酶通过氢化钠(NaBH4)处理达到稳定。该配合物应用于柱层析时,由于其不同的配合物稳定系数,允许分离各种脱氢酶。来自肝脏的乙醇脱氢酶、乳酸脱氢酶和腺苷酸激酶都与凝胶基质的adp类似物残基结合,因此可以通过不同的盐梯度分离。然而,酵母中的醇脱氢酶不会形成复合物,可以很容易地用磷酸盐缓冲液从色谱柱中洗脱出来。甘油醛-3磷酸和醛脱氢酶可以通过在缓冲液中加入NAD+或NADH来洗脱。还原的辅酶的不带电的1,4-二氢吡啶环与脱氢酶产生比氧化形式更稳定的配合物。
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引用次数: 1
Effects of membrane-acting drugs and aerobiosis on production of streptolysin S and nuclease in hemolytic streptococci. 膜作用药物和好氧作用对溶血性链球菌溶血素S和核酸酶产生的影响。
Pub Date : 1984-11-01 DOI: 10.1515/znc-1984-11-1222
A Taketo, Y Taketo

Yield of streptolysin S (SLS) in streptococcal culture was considerably reduced by procaine, dibucaine, atropine or chlorpromazine at concentrations which scarcely affected production of an extracellular nuclease as well as the bacterial growth. Cerulenin was also inhibitory to SLS formation, but its effect was more pronounced on the nuclease production. By aerobiosis, amount of SLS produced into culture supernatant was increased significantly, whereas yield of the nuclease was rather unaffected.

普鲁卡因、二布卡因、阿托品或氯丙嗪在几乎不影响胞外核酸酶的产生和细菌生长的浓度下,链球菌培养中溶链霉素S (SLS)的产量显著降低。蓝草素对SLS的形成也有抑制作用,但对核酸酶的产生作用更为明显。通过好氧作用,培养上清中产生的SLS数量显著增加,而核酸酶的产量则不受影响。
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引用次数: 0
High energy radiation-induced crosslinking of histone octamer complexes. 高能辐射诱导组蛋白八聚体复合物的交联。
Pub Date : 1984-11-01 DOI: 10.1515/znc-1984-11-1213
K J Deeg, L Katsikas, W Schnabel

Calf thymus histone octamer complexes were irradiated in the native state in N2O-saturated dilute aqueous solution (0.5 g/l, pH 9, [NaClO4] = 1 -4 mol/l) with 50 or 100 ns pulses of 16 MeV electrons or 60Co-gamma-rays. Time resolved light scattering measurements and optical absorption measurements yielded the following: the octamers underwent a volume contraction due to intra-complex-crosslinking induced by the attack of OH-radicals. Crosslinking proceeded to a certain extent via 2,2'-biphenol coupling as inferred from product analyses.

小牛胸腺组蛋白八聚体配合物在n2o饱和的稀水溶液(0.5 g/l, pH 9, [NaClO4] = 1 -4 mol/l)中以50或100 ns的16 MeV电子或60 co - γ射线脉冲照射。时间分辨光散射测量和光吸收测量得出如下结论:由于oh自由基的攻击导致配合物内部交联,八聚体发生了体积收缩。从产品分析推断,通过2,2'-双酚偶联,交联在一定程度上进行。
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引用次数: 3
Isolation of the products resulting from the reaction of cis and trans diaminedichloroplatinum [II] with DNA and chromatin on the Dowex 50 W column. 顺式和反式二胺二氯铂[II]与DNA和染色质反应产物在Dowex 50 W色谱柱上的分离。
R Oliński, Z Walter

The production of platinated derivatives of nucleic acid bases resulting from the reaction of cis and trans DDP with DNA and chromatin was studied. Bifunctional complex of guanine appeared to be the major product of the interaction of cis isomer with both DNA and chromatin, although other bifunctional adducts of A-Pt-G and A-Pt-A were also isolated. The main product of the interaction of trans DDP with DNA was a monofunctional adduct of guanine. Small amounts of the bifunctional complexes were also isolated. When ssDNA was incubated with trans DDP more bifunctional complexes appeared, what suggests that geometric constrains of double helix prevent formation of these complexes. Trans isomer reacts more easily with chromosomal proteins than cis DDP does. Therefore after the reaction of trans DDP with chromatin less platination occurs on DNA moieties.

研究了顺式和反式DDP与DNA和染色质反应生成核酸碱基铂化衍生物的工艺。鸟嘌呤双功能复合物似乎是顺式异构体与DNA和染色质相互作用的主要产物,尽管A-Pt-G和A-Pt-A的其他双功能加合物也被分离出来。反式DDP与DNA相互作用的主要产物是鸟嘌呤的单功能加合物。少量的双功能复合物也被分离出来。当ssDNA与反式DDP孵育时,出现了更多的双功能复合物,这表明双螺旋的几何约束阻止了这些复合物的形成。反式异构体比顺式DDP更容易与染色体蛋白发生反应。因此,在反式DDP与染色质反应后,DNA片段上发生较少的铂化。
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引用次数: 0
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Zeitschrift fur Naturforschung. Section C, Biosciences
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