Biological chemistry Hoppe-Seyler最新文献_第2页

Video-imaging micro-fluorometric assessment of luminal chloride/bicarbonate exchange activity in Madin-Darby canine kidney cells: influence of cell density, 4,4'-diisothiocyano-2,2'-disulfonic stilbene and acetazolamide. Madin-Darby犬肾细胞中氯/碳酸氢盐交换活性的视频成像微荧光评估:细胞密度、4,4'-二异硫氰基-2,2'-二磺酸二苯乙烯和乙酰唑胺的影响。

Biological chemistry Hoppe-Seyler Pub Date : 1996-05-01 DOI: 10.1515/bchm3.1996.377.5.275

S Ebner, M Marin-Grez

{"title":"Video-imaging micro-fluorometric assessment of luminal chloride/bicarbonate exchange activity in Madin-Darby canine kidney cells: influence of cell density, 4,4'-diisothiocyano-2,2'-disulfonic stilbene and acetazolamide.","authors":"S Ebner, M Marin-Grez","doi":"10.1515/bchm3.1996.377.5.275","DOIUrl":"https://doi.org/10.1515/bchm3.1996.377.5.275","url":null,"abstract":"To investigate whether or not MDCK cells may be used as a model for beta-intercalated cells, we studied: (1) The effect of luminal [Cl-]0 changes on pHi measured by video-imaging micro-fluorometry, (2) the influence of the inhibitor 4,4'-diisothiocyano-2,2'-disulfonic stilbene (DIDS) on anion-exchange activity, and (3) the effect of acetazolamide on intracellular pH-indicator (c-SNAFL-2) accumulation and anion-exchange activity. At least three different modes of fluorescence accumulation were found in confluent monolayers: cells with high, low or undetectable fluorescence. Highly fluorescent cells responded to a rise of [Cl-]0 (30-140 mM) with a proportional decrease of pHi (7.6-6.4). Acetazolamide (10(-4) M) completely blocked the acidifying effects of the increased [Cl-]0, indicating that HCO3- is the intracellular ion exchanged for extra-cellular Cl-. Acetazolamide caused a reduction of SNAFL-2 fluorescence suggesting that carbonic anhydrase activity contributes to indicator accumulation. The high DIDS concentration (50 microM) required to prevent intracellular acidification suggests that the exchanger involved is identical to that present in beta-intercalated cells. All cells of non-confluent monolayers were highly fluorescent and expressed Cl-/ HCO3(-)-exchanger activity. In conclusion, highly fluorescent MDCK cells in confluent monolayers have a luminal DIDS inhibitable, carbonic anhydrase dependent Cl-/HCO3(-)-exchanger, and may therefore be used as a model for beta-intercalated cells.","PeriodicalId":8963,"journal":{"name":"Biological chemistry Hoppe-Seyler","volume":"377 5","pages":"275-81"},"PeriodicalIF":0.0,"publicationDate":"1996-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/bchm3.1996.377.5.275","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19797210","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 3

Human colon carcinoma cells synthesize and secrete alpha 1-proteinase inhibitor. 人结肠癌细胞合成和分泌α 1蛋白酶抑制剂。

Biological chemistry Hoppe-Seyler Pub Date : 1996-05-01 DOI: 10.1515/bchm3.1996.377.5.301

D Keppler, M Markert, B Carnal, J Berdoz, J Bamat, B Sordat

{"title":"Human colon carcinoma cells synthesize and secrete alpha 1-proteinase inhibitor.","authors":"D Keppler, M Markert, B Carnal, J Berdoz, J Bamat, B Sordat","doi":"10.1515/bchm3.1996.377.5.301","DOIUrl":"https://doi.org/10.1515/bchm3.1996.377.5.301","url":null,"abstract":"Our previous results have shown that tumor cell-secreted procathepsin B can be activated by neutrophil elastase in vitro. In the present study, we addressed two questions: 1. Can neutrophil elastase be detected in human colon carcinomas, and 2. Does the co-culture of human colon carcinoma cells with neutrophils generate a cathepsin B-dependent pericellular proteolysis as assessed with radiolabeled laminin? We show that neutrophil elastase is present in colon carcinoma tissue and that its level is in good agreement with the degree of tissue infiltration by neutrophils. In co-culture experiments, elastase is released by neutrophils in a cell number dependent way, but no activation of tumor cell-secreted procathepsin B could be observed. In addition, the degradation of radiolabeled laminin by neutrophil proteinases was markedly decreased in the presence of tumor cells. These findings prompted us to search for a tumor cell-secreted elastase inhibitor. We show by enzyme activity measurements, gelatin-zymography, immunoblotting and RT-PCR that colon carcinoma cells synthesize and secrete alpha 1-proteinase inhibitor, a functional inhibitor of neutrophil elastase. The importance of this finding in the context of pericellular activation of tumor cell-secreted procathepsin B by neutrophil elastase is discussed.","PeriodicalId":8963,"journal":{"name":"Biological chemistry Hoppe-Seyler","volume":"377 5","pages":"301-11"},"PeriodicalIF":0.0,"publicationDate":"1996-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/bchm3.1996.377.5.301","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19797213","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 25

GSH transporters: molecular characterization and role in GSH homeostasis. 谷胱甘肽转运体:分子表征及其在谷胱甘肽稳态中的作用。

Biological chemistry Hoppe-Seyler Pub Date : 1996-05-01 DOI: 10.1515/bchm3.1996.377.5.267

N Kaplowitz, J C Fernández-Checa, R Kannan, C Garcia-Ruiz, M Ookhtens, J R Yi

引用次数: 95

Isolation and characterization of sialate lyase from pig kidney. 猪肾中唾液酸裂解酶的分离与鉴定。

Biological chemistry Hoppe-Seyler Pub Date : 1996-05-01 DOI: 10.1515/bchm3.1996.377.5.293

R Schauer, M Wember

{"title":"Isolation and characterization of sialate lyase from pig kidney.","authors":"R Schauer, M Wember","doi":"10.1515/bchm3.1996.377.5.293","DOIUrl":"https://doi.org/10.1515/bchm3.1996.377.5.293","url":null,"abstract":"Sialate lyase (sialate aldolase; systematic name N-acetylneuraminate pyruvate-lyase, EC 4.1.3.3) was isolated as soluble enzyme from pig kidney and purified 630-fold using a heating step, gel filtration, and chromatography on immobilized neuraminic acid beta-methyl glycoside in 14% yield to apparent homogeneity as tested by SDS-gel electrophoresis. The molecular mass is 58 kDa and the pH-optimum is at pH 7.2. Kinetic parameters were determined with N-acetyl-neuraminic acid as substrate: Km 3.7 mM and Vmax 37.1 mU. The lyase cleaves only free sialic acids with relative rates of 100% for N-acetylneuraminic acid, 55% for N-glycolylneuraminic acid and 32% for N-acetyl-9-O-acetylneuraminic acid, whereas N-acetyl-4-O-acetylneuraminic acid or 2-deoxy-2,3-didehydro-N-acetylneuraminic acid are not substrates. Enzyme activity was inhibited with p-chloromercuribenzoate, o-phenanthroline, cyanide, 5-diazonium-1-H-tetrazole, 5,5'-dithiobis(2-nitrobenzoic acid), diethylpyro-carbonate, and Rose Bengal in the presence of light and O2. Reduction with sodium borohydride in the presence of N-acetylneuraminic acid or pyruvate resulted in irreversible inhibition of enzyme activity. The inhibition experiments suggest the involvement of histidine, lysine and SH-residues in enzyme catalysis. Thus, this mammalian lyase most probably belongs to the Class I aldolases, and has properties similar to the same enzyme from Clostridium perfringens and is active with the alpha-form of N-acetylneuraminic acid.","PeriodicalId":8963,"journal":{"name":"Biological chemistry Hoppe-Seyler","volume":"377 5","pages":"293-9"},"PeriodicalIF":0.0,"publicationDate":"1996-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/bchm3.1996.377.5.293","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19797212","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 20

Purification and characterisation of the pyruvate decarboxylase from a haploid strain of Saccharomyces cerevisiae. 酿酒酵母菌单倍体丙酮酸脱羧酶的纯化与特性研究。

Biological chemistry Hoppe-Seyler Pub Date : 1996-05-01 DOI: 10.1515/bchm3.1996.377.5.313

M Killenberg-Jabs, S König, S Hohmann, G Hübner

引用次数: 16

Species specificity of thimet oligopeptidase (EC 3.4.24.15). 硫柳寡肽酶的物种特异性(EC 3.4.24.15)。

Biological chemistry Hoppe-Seyler Pub Date : 1996-05-01 DOI: 10.1515/bchm3.1996.377.5.283

M A Hayashi, M D Gomes, N A Rebouças, B L Fernandes, E S Ferro, A C de Camargo

{"title":"Species specificity of thimet oligopeptidase (EC 3.4.24.15).","authors":"M A Hayashi, M D Gomes, N A Rebouças, B L Fernandes, E S Ferro, A C de Camargo","doi":"10.1515/bchm3.1996.377.5.283","DOIUrl":"https://doi.org/10.1515/bchm3.1996.377.5.283","url":null,"abstract":"The recombinant rat testes metallo-endooligopeptidase (EC 3.4.24.15) and the rabbit brain endooligopeptidase A (formerly EC 3.4.22.19) were compared, side-by-side, in view of their striking similarities in both the physicochemical features and the specificities for oligopeptides. Concerning the tissue distribution in rat and rabbit, no relation between the levels of enzyme activity in cytosol and the levels of metallo-endooligopeptidase 24.15 mRNA could be established. The results suggest that the predominant neuropeptide-metabolizing activity attributed to the metallo-endooligopeptidase 24.15 is performed by, at least, two distinct cytosolic enzymes, one predominant in rat testes and the other in rabbit brain and testes, and possibly also in rat brain. Both enzymes are activated by dithiothreitol and irreversibly inhibited by a SH-affinity labeling dynorphin-related compound, but they are not inhibited by EDTA in a concentration dependent manner. Both enzymes exhibit the same specificity toward several bioactive peptides, except for LH-RH and substance P, which are only hydrolysed by the rat testes enzyme. Taken together, these results lead us to conclude that it is unlikely that the recombinant rat testes metallo-endooligopeptidase 24.15 and the rabbit brain endooligopeptidase A are the same molecule although they might belong to the same family of oligopeptidases.","PeriodicalId":8963,"journal":{"name":"Biological chemistry Hoppe-Seyler","volume":"377 5","pages":"283-91"},"PeriodicalIF":0.0,"publicationDate":"1996-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/bchm3.1996.377.5.283","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19797211","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 13

Dramatic changes in the ratio of homologous recombination to nonhomologous DNA-end joining in oocytes and early embryos of Xenopus laevis. 非洲爪蟾卵母细胞和早期胚胎中同源重组与非同源dna末端连接比率的显著变化。

Biological chemistry Hoppe-Seyler Pub Date : 1996-04-01 DOI: 10.1515/bchm3.1996.377.4.239

M Hagmann, K Adlkofer, P Pfeiffer, R Bruggmann, O Georgiev, D Rungger, W Schaffner

{"title":"Dramatic changes in the ratio of homologous recombination to nonhomologous DNA-end joining in oocytes and early embryos of Xenopus laevis.","authors":"M Hagmann, K Adlkofer, P Pfeiffer, R Bruggmann, O Georgiev, D Rungger, W Schaffner","doi":"10.1515/bchm3.1996.377.4.239","DOIUrl":"https://doi.org/10.1515/bchm3.1996.377.4.239","url":null,"abstract":"We have developed a versatile plasmid vector (pReco-sigma) for recombination studies. When linearized and introduced into the cells of interest, pReco-sigma allows the simultaneous determination of the relative frequencies of homologous recombination versus nonhomologous DNA-end joining (also termed end-to-end joining), the latter an example of illegitimate recombination processes. As a system we made use of stage VI oocytes and fertilized eggs of the African clawed frog Xenopus laevis, which were previously described to support homologous recombination and DNA-end joining, respectively. Extending these earlier findings, we show that oocytes yield > 80% of the homologously recombined product, whereas in eggs a highly efficient DNA-end joining activity predominates (> 95%). Both reactions, homologous recombination and DNA-end joining, are shown to occur quickly, with the majority of the respective products being formed within the first 20 minutes of incubation under optimal conditions. In fertilized eggs, up to 50% of all injected linear DNA molecules are recircularized by DNA-end joining. With high amounts of injected DNA per fertilized egg, DNA-end joining is reduced, presumably due to competition for essential factors, and homologous recombination becomes readily detectable. As there is a sequence of rapid cleavage divisions after fertilization of the egg, the fast and highly efficient DNA-end joining, even though it is error-prone at the junction site, seems to be best suited to cope with DNA double-strand breaks that might occur in the genome during early embryogenesis. On the other hand, the long-lived oocytes seem to repair DNA double-strand breaks via homologous recombination. This latter property may be exploited both in Xenopus and in other organisms to achieve homologous integration of exogenous DNA into germ cells for gene targeting.","PeriodicalId":8963,"journal":{"name":"Biological chemistry Hoppe-Seyler","volume":"377 4","pages":"239-50"},"PeriodicalIF":0.0,"publicationDate":"1996-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/bchm3.1996.377.4.239","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19710701","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 40

Generation and characterization of inducible nitric oxide synthase deficient macrophage cell lines. 诱导型一氧化氮合酶缺陷巨噬细胞系的产生及特性研究。

Biological chemistry Hoppe-Seyler Pub Date : 1996-04-01 DOI: 10.1515/bchm3.1996.377.4.227

H Rothe, G Bosse, H G Fischer, H Kolb

{"title":"Generation and characterization of inducible nitric oxide synthase deficient macrophage cell lines.","authors":"H Rothe, G Bosse, H G Fischer, H Kolb","doi":"10.1515/bchm3.1996.377.4.227","DOIUrl":"https://doi.org/10.1515/bchm3.1996.377.4.227","url":null,"abstract":"Stable inducible nitric oxide synthase deficient mouse macrophage cell lines were generated by the antisense technology. A 666 bp fragment of a mouse inducible nitric oxide synthase cDNA was cloned in antisense orientation into a mammalian expression vector behind the CMV promoter. This construct was transfected into J774.1A cells, a mouse macrophage cell line. The inducible nitric oxide synthase antisense lines showed up to 84% reduction of nitric oxide production in response to lipopolysaccharide stimulation and 66% reduction of nitric oxide production in response to interferon-gamma and a combination of interferon-gamma and lipopolysaccharide stimulation. The deficiency in inducible nitric oxide synthase expression had no impact on lipopolysaccharide induced tumor necrosis factor alpha and interleukin-1 secretion. The stable and specific inhibition of inducible nitric oxide synthase expression by antisense DNA vectors allows a direct analysis of contribution of inducible nitric oxide synthase activity to macrophage regulatory and immune defence functions.","PeriodicalId":8963,"journal":{"name":"Biological chemistry Hoppe-Seyler","volume":"377 4","pages":"227-31"},"PeriodicalIF":0.0,"publicationDate":"1996-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/bchm3.1996.377.4.227","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19710698","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 8

The rps4-gene is encoded upstream of the nad2-gene in Arabidopsis mitochondria. rps4基因编码在拟南芥线粒体nad2基因的上游。

Biological chemistry Hoppe-Seyler Pub Date : 1996-04-01 DOI: 10.1515/bchm3.1996.377.4.251

B Lippok, A Brennicke, M Unseld

引用次数: 16

The structures of the bovine and porcine proacrosin genes and their conservation among mammals. 牛和猪的促红素基因结构及其在哺乳动物中的保存。

Biological chemistry Hoppe-Seyler Pub Date : 1996-04-01

I M Adham, H Kremling, S Nieter, S Zimmermann, M Hummel, U Schroeter, W Engel

引用次数: 0