Biological chemistry Hoppe-Seyler最新文献_第10页

Quantification of intracellular cathepsin activities in human lung tumor cell lines by flow cytometry. 流式细胞术定量人肺肿瘤细胞系细胞内组织蛋白酶活性。

Biological chemistry Hoppe-Seyler Pub Date : 1995-07-01 DOI: 10.1515/bchm3.1995.376.7.407

B Ulbricht, E Spiess, R Schwartz-Albiez, W Ebert

{"title":"Quantification of intracellular cathepsin activities in human lung tumor cell lines by flow cytometry.","authors":"B Ulbricht, E Spiess, R Schwartz-Albiez, W Ebert","doi":"10.1515/bchm3.1995.376.7.407","DOIUrl":"https://doi.org/10.1515/bchm3.1995.376.7.407","url":null,"abstract":"The cysteine proteases cathepsin B and cathepsin L are very likely involved in invasive processes of normal and malignant cells, they become relevant for a number of diseases and are possibly prognostic markers for the outcome of human lung cancer. Therefore, we have determined activities of these related enzymes in cells and in cell extracts of human lung carcinoma cell lines of different cathepsin composition by flow cytometry and by spectrophotometry, respectively. To this end we applied the synthetic dipeptidyl substrates benzoxycarbonyl-arginyl-arginine- and benzoxycarbonyl-phenyl-arginine- coupled to 4-methoxy-beta-naphthylamide, aminomethyl-coumarine or rhodamine R110. The apparent enzymatic activities were differentially defined by protease inhibitors, particularly E-64 and CA-074. Independent of the dipeptidyl-composition more than 99 per cent of the apparent activity was due to cathepsin B when 4-methoxy-beta-naphthylamide or aminomethylcoumarine were the leaving groups. The 4-methoxy-beta-naphthylamide precipitate used for detection of cell associated activities revealed a wide spectrum of excitation to fluorescence thwarting the application of other possible fluorescent tags. Therefore, its application is restricted to uniparametric fluorescence investigations. Both dipeptidylgroups coupled to rhodamine R110 were promiscuous: only 25 to 30% of the apparent activity were due to cathepsin B; the predominant activity came from cathepsin L, irrespective whether intracellular or activities of cellular extracts were analyzed. However, rhodamine R110-coupled substrates open the way for multiparametric fluorescent analysis of cathepsins B and L containing cells if appropriate inhibitors for specification of the enzymatic activities are additionally applied. In very contrast to 4-methoxy-beta-naphthylamide, which causes irreparable damage to the cells, the rhodamine substrates permit studies with living cells and live cell sorting.","PeriodicalId":8963,"journal":{"name":"Biological chemistry Hoppe-Seyler","volume":"376 7","pages":"407-14"},"PeriodicalIF":0.0,"publicationDate":"1995-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/bchm3.1995.376.7.407","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18581764","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

In vitro fibril formation from alpha 1-antitrypsin-derived C-terminal peptides. α 1-抗胰蛋白酶衍生的c端肽在体外形成纤维。

Biological chemistry Hoppe-Seyler Pub Date : 1995-07-01 DOI: 10.1515/bchm3.1995.376.7.415

S Janciauskiene, E Carlemalm, S Eriksson

{"title":"In vitro fibril formation from alpha 1-antitrypsin-derived C-terminal peptides.","authors":"S Janciauskiene, E Carlemalm, S Eriksson","doi":"10.1515/bchm3.1995.376.7.415","DOIUrl":"https://doi.org/10.1515/bchm3.1995.376.7.415","url":null,"abstract":"Fragments from various proteolytically degraded precursor proteins can form beta-amyloid fibrils. We studied, by electron microscopy and quantitative Congo red binding, the ability of three synthetic peptides, corresponding to residues 359-374 (C-36), 370-374 (C-5) and 375-394 (C-20) from the C-terminal part of alpha 1-antitrypsin (AAT) to form beta-amyloid fibrils in vitro. The peptides C-36 and C-5 had a pronounced tendency to form fibrils. C-20 lacked this property, suggesting that residues 359-375 and/or 370-374 are most critical for fibril formation. Native AAT added to peptide 125I-C-36 could bind and form complexes with the peptide, resulting in inhibition of amyloid fibril formation. Moreover, native AAT added to preformed fibrils induced disaggregation of fibrillar structures. The structural rearrangements of AAT that occurred during this 'autointeraction' included polymerization of the serpin, and an increase of its thermal stability. Also, following interaction, an increase (20-40%) of AAT's antielastase activity was noted. The demonstration of an in vitro beta-amyloid fibril formation from the AAT derived C-terminal peptides C-36 and C-5 and its regulation by the intact AAT molecule may have important in vivo implications.","PeriodicalId":8963,"journal":{"name":"Biological chemistry Hoppe-Seyler","volume":"376 7","pages":"415-23"},"PeriodicalIF":0.0,"publicationDate":"1995-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/bchm3.1995.376.7.415","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18581765","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

Amino acid sequence of alpha- and beta-polypeptide chains of turkey (Meleagris gallopavo) hemoglobin. 火鸡血红蛋白α -和β -多肽链氨基酸序列。

Biological chemistry Hoppe-Seyler Pub Date : 1995-07-01

Y Eguchi, T Ikehara, S Kayo, T Eguchi, H Takei

引用次数: 0

Identification of an alanine aminopeptidase in human maternal serum as a membrane-bound aminopeptidase N. 人母血清丙氨酸氨基肽酶的膜结合氨基肽酶N的鉴定。

Biological chemistry Hoppe-Seyler Pub Date : 1995-07-01 DOI: 10.1515/bchm3.1995.376.7.397

Y Watanabe, S Iwaki-Egawa, H Mizukoshi, Y Fujimoto

引用次数: 18

Molecular cloning and characterization of the bovine and porcine outer dense fibers cDNA and organization of the bovine gene. 牛、猪外密纤维cDNA的分子克隆与鉴定及牛基因的组织。

Biological chemistry Hoppe-Seyler Pub Date : 1995-07-01 DOI: 10.1515/bchm3.1995.376.7.431

Y Kim, I M Adham, T Haack, H Kremling, W Engel

{"title":"Molecular cloning and characterization of the bovine and porcine outer dense fibers cDNA and organization of the bovine gene.","authors":"Y Kim, I M Adham, T Haack, H Kremling, W Engel","doi":"10.1515/bchm3.1995.376.7.431","DOIUrl":"https://doi.org/10.1515/bchm3.1995.376.7.431","url":null,"abstract":"Outer dense fibers (ODF) or accessory fibers are filamentous structures of the sperm tail of many eumetozoan organisms endowed with internal fecundation. The bovine and porcine cDNA of an outer dense fiber protein was cloned, sequenced and compared to the previously characterized human and rat cDNA sequences. The coding sequences and the 5' and 3' untranslated regions of the ODF cDNAs are highly conserved. A comparison of the bovine, porcine, human and rat ODF protein sequences revealed that the protein displays a high degree of similarity, ranging from 87% to 98%. The ODF protein is rich in cysteine and contains the C.X.P. repeat at the C-terminal which is different in number among mammalian species. All the 27 cysteine residues in the ODF sequence except those in the C.X.P. repeat are conserved in the four species. We report here also the organization of the bovine ODF gene which is similar to that of human and rat. The transcription start site in the bovine ODF gene is localized 98 bp upstream of the translation start site. Alignment of the 5' flanking region of bovine ODF with the rat gene reveals that the first 130 nucleotides upstream of the transcription start site exhibit an overall sequence similarity of 83%. This conserved region contains a TATA-like box (TTTAAA) and binding sites for AFT/CREB and EGR-1 transcription factors.","PeriodicalId":8963,"journal":{"name":"Biological chemistry Hoppe-Seyler","volume":"376 7","pages":"431-5"},"PeriodicalIF":0.0,"publicationDate":"1995-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/bchm3.1995.376.7.431","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18581767","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: 17

Human cathepsin O2, a novel cysteine protease highly expressed in osteoclastomas and ovary molecular cloning, sequencing and tissue distribution. 破骨细胞瘤和卵巢中高表达的新型半胱氨酸蛋白酶组织蛋白酶O2的分子克隆、测序和组织分布。

Biological chemistry Hoppe-Seyler Pub Date : 1995-06-01 DOI: 10.1515/bchm3.1995.376.6.379

D Brömme, K Okamoto

引用次数: 232

Visualization of tissue kallikrein in human breast carcinoma by two-dimensional western blotting and immunohistochemistry. 二维western blotting和免疫组织化学技术在人乳腺癌组织中的可视化研究。

Biological chemistry Hoppe-Seyler Pub Date : 1995-06-01 DOI: 10.1515/bchm3.1995.376.6.365

A Hermann, P Buchinger, J Rehbock

引用次数: 22

Cytokine regulation of matrix metalloproteinase activity and its regulatory dysfunction in disease. 细胞因子对基质金属蛋白酶活性的调节及其在疾病中的调节功能障碍。

Biological chemistry Hoppe-Seyler Pub Date : 1995-06-01

C Ries, P E Petrides

{"title":"Cytokine regulation of matrix metalloproteinase activity and its regulatory dysfunction in disease.","authors":"C Ries, P E Petrides","doi":"","DOIUrl":"","url":null,"abstract":"Matrix metalloproteinases (MMPs) represent a family of structurally and functionally related enzymes responsible for the proteolytic degradation of extracellular matrix (ECM) components such as basement membrane or interstitial stroma. MMPs are important participants of normal tissue remodeling. Due to their potential hazardous effects MMPs are highly regulated at different levels. At the transcriptional level, MMP expression is precisely controlled by various cytokines acting through positive or negative regulatory elements of its genes. Moreover, MMP activity is post-transcriptionally regulated by proteolytic activation of the latent proenzymes and by interaction with specific tissue inhibitors of metalloproteinases (TIMPs). Expression and secretion of both MMP activating enzymes and TIMPs are also influenced by cytokines. Dysregulation of MMP production and activation may cause altered extracellular proteolysis that is associated with a number of diseases such as rheumatoid arthritis and tumor metastasis. Thus, the molecular analysis of the regulatory mechanisms of gene expression and activity of MMPs and their inhibitors is essential for understanding the complex scenario of tissue remodeling and ECM degradation under both normal and pathological conditions.","PeriodicalId":8963,"journal":{"name":"Biological chemistry Hoppe-Seyler","volume":"376 6","pages":"345-55"},"PeriodicalIF":0.0,"publicationDate":"1995-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18583549","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: 0

Purification and N-terminal amino-acid sequence analysis of rabbit neutrophil cathepsin G. 兔中性粒细胞组织蛋白酶G的纯化及n端氨基酸序列分析。

Biological chemistry Hoppe-Seyler Pub Date : 1995-06-01 DOI: 10.1515/bchm3.1995.376.6.371

E Cavarra, A Santucci, G Lungarella

{"title":"Purification and N-terminal amino-acid sequence analysis of rabbit neutrophil cathepsin G.","authors":"E Cavarra, A Santucci, G Lungarella","doi":"10.1515/bchm3.1995.376.6.371","DOIUrl":"https://doi.org/10.1515/bchm3.1995.376.6.371","url":null,"abstract":"Cathepsin G was isolated from granules of rabbit bloodstream leukocytes and purified to apparent homogeneity by a multi-step procedure consisting of ammonium sulphate precipitation, affinity chromatography on elastin-Sepharose, and finally by ion-exchange chromatography on a CM-52 column. The molecular weight of the enzyme, as determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), was 27,000. The first 24 N-terminal amino-acids were determined and showed 96%, 92% and 79% identity respectively to those of human, dog and rat cathepsin G. Despite the difference in the total amino-acid composition of cathepsin G between rabbit and other mammalian species, close similarities have been found in their substrate specificity and inhibition profile. The kcat/Km values of rabbit cathepsin G with Suc-Ala2-Pro-Phe-NA and Suc-Ala2-Pro-Leu-NA are quite similar to those reported for human cathepsin G under the same conditions. The inhibition profile of the isolated enzyme indicates that cathepsin G from rabbits, like that from other mammalians species belongs to the group of serine proteinases. Finally, like human cathepsin G, catalytically active rabbit enzyme is able to induce platelet aggregation.","PeriodicalId":8963,"journal":{"name":"Biological chemistry Hoppe-Seyler","volume":"376 6","pages":"371-7"},"PeriodicalIF":0.0,"publicationDate":"1995-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1515/bchm3.1995.376.6.371","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18583552","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: 1

Expression of full-length human procathepsin L cDNA in Escherichia coli and refolding of the expression product. 全长人血凝素原L cDNA在大肠杆菌中的表达及表达产物的重折叠。

Biological chemistry Hoppe-Seyler Pub Date : 1995-06-01 DOI: 10.1515/bchm3.1995.376.6.385

M Dolinar, D B Maganja, V Turk

引用次数: 36