Dosages protéiques et profils d''activités enzymatiques comparés chez les deux espèces de babouins Papio anubis et P. cynocephalus

Biochemical levels of serum proteins and enzymes were investigated in two populations of the baboons Papio anubis and P. cynocephalus. The mean average levels of proteins or enzyme activity are the same in the two species. The only difference concerns the level of an inhibitor of pseudocholinesterase in a few individuals of Papio anubis.     

The glutathione system. I. Synthesis, transport, glutathione transferases, glutathione peroxidases

During the last 10–15 years significant progress has been achieved in all directions of studies of the glutathione system. A series of new enzymes involved into metabolism of glutathione has been discovered. Many of these enzymes are polyfunctional and their new activities have been recognized. The enzymes interact with hormones and signal transduction systems. Significant progress has been achieved in the studies of intracellular, intercellular and inter-organ transport. The important achievement is employment of not only selective compounds-analyzers but also gene engineering methods for identification of new functions.     

Characterisation of a secretory antigen from Toxoplasma gondii and its role in circulating antigen production

Hughes H. P. A. and van Knapen F. 1982. Characteristics of a secretory antigen from Toxoplasma gondii and its role in circulating antigen production. International Journal for Parasitology12: 433–437. In vitro culture of RH Toxoplasma gondii in HEp2 cells was found to yield an antigen, of mol. wt. 324,000 dallons, which is one of the components of circulating antigen (CAg). Hydrophobic interaction electrophoresis of ¹²⁵I labelled solubilised parasites has shown that this antigen, in common with the other CAg component, is of intracellular origin. Cyclophosphamide had no effect on either parasite proliferation or on secretion of antigen in vitro. Although immune lysis appears to be the major pathway of CAg release in vivo, secretion by the parasite may be important in the expression of CAg in serum and body fluids immediately following infection.     

Additional studies of sheep haemopexin: genetic control,frequencies and postnatal development*

Summary. This study presents evidence that sheep haemopexin phenotypes are genetically controlled by three alleles, Hpx^A, Hpx^B1 and Hpx^B2, of a single autosomal locus. Frequencies of two alleles, Hpx^A and Hpx^B (Hpx^B encompasses two isoalleles, Hpx^B1 and Hpx^B2), were studied in eight sheep breeds in Czechoslovakia. The frequency of the HpxA allele was highest (ranging from 0.81 in Merino to 1.0 in East Friesian sheep). Qualitative and quantitative changes in haemopexin during postnatal development were studied by starch gel electrophoresis and rocket immunoelectrophoresis respectively. In electrophoresis, 1- or 2-day-old lambs had two very weak zones corresponding in mobility to two slower zones of adult animals. Later, the third more anodic zone appeared and gradually increased in intensity. In 1-month-old lambs the patterns were practically identical with those of adult animals. Using rocket immunoelectrophoresis, the level of haemopexin shortly after birth was practically zero. It rose sharply till the sixth day of life; then the level continued to rise slowly till about 1 month of age. The mean haemopexin level in adult sheep was 64.5 ± 18.26 (SD) mg/100ml serum, ranging from 30.5 to 116.5 mg/100ml.     

酵母麦角固醇生物合成及其基因调控的研究

麦角固醇是真菌细胞膜中的重要组成成分,是维生素D2的合成前体,也是一种重要医药材料。近年来,麦角固醇生物合成途径已经被弄清,其基因调控也获得了初步的进展。就麦角固醇的生物合成途径、酶在细胞内的分布及其基因调控进行综述。     

Rocket and crossed immunoelectrophoresis of proteins solubilized with sodium dodecyl sulfate

A method for the immunoelectrophoretic analysis of both hydrophilic and hydrophobic proteins from whole-cell extracts solubilized with 2% (w/v) sodium dodecyl sulfate (SDS) is described. For rocket immunoelectrophoresis, Triton X-100 is added to the sample before electrophoresis to sequester non-protein-bound SDS, and polyethylene glycol (PEG) is added to the antibody gel to enhance precipitin formation. With the optimal ratio of Triton X-100 to PEG, the quantitative determination of 5 ng of protein is possible. The SDS-solubilized sample can also be analyzed by crossed immunoelectrophoresis using SDS-polyacrylamide gels in the first dimension and antibody-containing agarose gels in the second. The best results are obtained when intermediate gels without nonionic detergents are used and when ionic detergents are omitted from the cathodal gel. Precipitin peaks of high quality, reproducibility, and without artifacts are obtained using antibody concentrations 5- to 50-fold lower than with other crossed-immunoelectrophoresis procedures.     

Evolutionary analysis of a novel zinc ribbon in the N-terminal region of threonine synthase

Threonine synthase (TS) catalyzes the terminal reaction in the biosynthetic pathway of threonine and requires pyridoxal phosphate as a cofactor. TSs share a common catalytic domain with other fold type II PALP dependent enzymes. TSs are broadly grouped into two classes based on their sequence, quaternary structure, and enzyme regulation. We report the presence of a novel zinc ribbon domain in the N-terminal region preceding the catalytic core in TS. The zinc ribbon domain is present in TSs belonging to both classes. Our sequence analysis reveals that archaeal TSs possess all zinc chelating residues to bind a metal ion that are lacking in the structurally characterized homologs. Phylogenetic analysis suggests that TSs with an N-terminal zinc ribbon likely represents the ancestral state of the enzyme while TSs without a zinc ribbon must have diverged later in specific lineages. The zinc ribbon and its N- and C-terminal extensions are important for enzyme stability, activity and regulation. It is likely that the zinc ribbon domain is involved in higher order oligomerization or mediating interactions with other biomolecules leading to formation of larger metabolic complexes.     

Purification,characterization, and synthesis of vitellin from the cabbage butterfly Pieris rapae L.

Vitellin from the cabbage butterfly Pieris rapae L. was purified and characterized by electrophoresis. Vitellin from P. rapae is a phosphorylated glycolipoprotein of 380,000 ± 10,000 molecular weight as determined by nondenaturing polyacrylamide gel electrophoresis. Two subunits with an M_r of 150,000 and 40,000 were obtained from vitellin. The native molecule is thought to be a tetramer composed of two molecules of each of these subunits. The isoelectric point, as determined by isoelectric focusing on polyacrylamide gels, is 6.10. Vitellin and vitellogenin were indistinguishable by immunological methods such as double diffusion and tandem-crossed immunoelectrophoresis. Vitellogenin from the hemolymph and vitellin from the ovary were quantified by rocket immunoelectrophoresis. Vitellogenin and vitellin were first detected in 6-day-old pupae, and their levels increased continuously during ovarian development. Vitellogenin synthesis by the fat body in 4-day-old female pupae could be induced by juvenile hormone I.     

Immunological properties of entomocidal protein of Bacillus thuringiensis and its insecticidal activity

The immunological properties of the proteinaceous component of the parasporal crystal (δ-endotoxin) of Bacillus thuringiensis var. kurstaki were analyzed by rocket immunoelectrophoresis. Two antisera, one against the k-l-type crystal containing two components, and the other against the k-73-type crystal containing one component, were made in rabbits. The antigens consisting of purified and dissociated crystals were run in electrophoresis with these two antisera. The ratio between the two peak heights of precipitin lines, which were formed by the dissociated crystal of one B. thuringiensis isolate in two antisera, was compared with the ratios of other isolates under identical conditions. The difference in the ratio reflected a difference in the structure of the crystal component and correlated closely with the insecticidal activity spectrum. This method can be used to evaluate a newly isolated B. thuringiensis, and it can further differentiate the isolates which have been classified as one serotype.     

A common-immunogenic Vibrio outer membrane protein

Abstract The presence of antibodies in rabbit antisera to cell envelope proteins of Vibrio cholerae has been examined using a two-dimensional system, in which the cell envelopes are electrophoresed in sodium dodecyl sulfate (SDS) in polyacrylamide gels in the first dimension, and in agarose containing antibodies in the second. The results show that a 25-kDa protein is markedly immunogenic and appears to be common to Vibrio strains; it is not present in a number of other organisms. This 25-kDa protein is an outer membrane protein as judged by sucrose gradient centrifugation and it is accessible to iodination by lactoperoxidase, suggesting that it is exposed on the cell surface.     

Comparative methods of detecting HbsAg in chronic glomerulonephritis patients in Nigeria

Abstract A comparative evaluation of the enzyme-linked immunosorbent assay (Elisa), passive haemagglutination (PHA) and counterimmunoelectrophoresis (CIE) methods were carried out. Approx. 6% of control samples were positive with the Elisa assay, while the values for Behring Institute (BI) passive haemagglutination, Wellcome Research Laboratory (WRL) passive haemagglutination and CIE were 2.7%, 2.2% and 3.3%, respectively. The above data contrast with values of 21.3%, 10.6, 10.3 and 12.3 obtained in sera from chronic glomerulonephritis patients.
Our observation suggests that HbsAg may be associated with chronic glomerulonephritis.     

Microscale purification of proteins by line immunoelectrophoresis: Application of the technique in protein biogenesis studies

A small-scale version of line immunoelectrophoresis in combination with immunoprecipitate excision is describeb as a rapid convenient technique to purify proteins on a micro scale in biogenesis studies. In the purification of pig small intestinal microvillar enzymes the method was found to be capable of a quantitative purification and to result in a higher state of purity than an isolation procedure using protein A-Sepharose. Since the method furthermore allows a simultaneous purification of several different protein antigens from the sample, it may be of interest as an alternative method to other procedures in the purification of proteins on a micro scale.     

Long term regulation of glycogen metabolizing enzymes by insulin in H4 hepatoma cells

Insulin alone at concentrations of less than 1 to 5 uU/ml increased the enzyme activities of glycogen synthase, synthase phosphatase, phosphorylase, and phosphorylase phosphatase in hepatoma H4 cells in culture in the presence and absence of serum. Increase in total and active forms of glycogen synthase and phosphorylase were observed. Cycloheximide blocked the action of insulin on glycogen synthase, glycogen synthase phosphatase and phosphorylase phosphatase. The enzymes with the exception of glycogen synthase phosphatase were expressed with greater hormonal sensitivity in the absence as compared to the presence of serum in terms of hormone concentration required and or time of onset.These results demonstrate that these glycogen metabolizing enzymes are under long term control by insulin, with glycogen synthase being the most sensitive of the enzymes studied to the action of the hormone.Supported by grants from NIH AM 14334 and AM 22125 (University of Virginia Diabetes Research and Training Center) and by a grant from Lilly Research Lab, and the March of Dimes     

Ca(NO3)2对NaCl胁迫下木麻黄扦插苗生理特征的调控

用不同浓度 Ca(NO3) 2 · 4 H2 O(0 .7、1.4、2 .1g Ca2 / kg土 )对 1年生的处在两种 Na Cl胁迫 (10和 2 0 g/ kg)处理下的木麻黄扦插苗进行化学调控 ,研究硝酸钙盐加氯化钠处理的木麻黄幼苗的生长量、木麻黄幼苗抗氧化酶系统活性和渗透调节物质的含量的变化 ,研究结果表明 :中度 Na Cl胁迫加硝酸钙处理下的木麻黄扦插苗的可溶性蛋白质含量增加 ,MDA含量降低说明膜脂过氧化作用减轻 ,而且抗氧化酶活性 (SOD、POD)之间协调变化有利于提高清除自由基的速率 ,中度盐胁迫下钙盐可以促进木麻黄体内脯氨酸的积累 ;但重度 Na Cl胁迫下钙盐对木麻黄的调控作用不显著 ,重度盐胁迫下钙盐反而降低木麻黄 SOD、POD活性和脯氨酸的含量 ,减弱了抗氧化酶系统对活性氧的清除作用 ,同时高浓度钙盐还会加重 Na Cl胁迫对木麻黄幼苗的损伤 ,这说明适量的钙盐有利于木麻黄幼苗抵抗盐胁迫能力的提高 ,而高浓度钙盐则可能会加重盐胁迫     

Molecular Polymorphism and Phylogenetic Relationship in some Meloidogyne spp.: Application to the Taxonomy of Meloidogyne

Proteins and various isozymes were investigated by direct analysis of single specimens in order to check molecular genetic variability, which is not rare in Meloidogyne species in spite of parthenogenetic reproduction. Variability was found in esterases, ocglycerophosphate, malate dehydrogenases, and some other proteins. Other loci appear monomorphic in the genus (for example, catalase), Distinct pools of genes are in a relative accordance with the common described species. Characteristic electrophoretograms are given for M. arenaria, M. javanica, M. incognita, M. hapla, and M. naasi, and it appears that nonspecific esterases are a useful tool supplementing morphology for specific characterization. Because the biochemical evidence is less subjective than the morphological, we believe it is more reliable.     

植物挥发性物质代谢的遗传修饰

植物在生命周期里能合成释放多种多样的挥发性化合物,这些物质在植物生长发育和代谢调控,植物抵御病虫侵害,以及植物与环境信息交流中行使重要功能。在介绍植物挥发性化合物的生理功能、合成途径调控和商业应用的基础上,重点论述应用基因工程调控植物挥发性物质合成的技术策略和研究进展,并讨论了植物挥发性物质遗传修饰存在的问题和发展前景。