Current chemical genomics最新文献_第2页

A homogenous luminescence assay reveals novel inhibitors for giardia lamblia carbamate kinase. 一种同质发光试验揭示了新的兰第鞭毛虫氨基甲酸酯激酶抑制剂。

Current chemical genomics Pub Date : 2012-01-01 Epub Date: 2012-12-31 DOI: 10.2174/1875397301206010093

Catherine Z Chen, Noel Southall, Andrey Galkin, Kap Lim, Juan J Marugan, Liudmila Kulakova, Paul Shinn, Danielle van Leer, Wei Zheng, Osnat Herzberg

{"title":"A homogenous luminescence assay reveals novel inhibitors for giardia lamblia carbamate kinase.","authors":"Catherine Z Chen, Noel Southall, Andrey Galkin, Kap Lim, Juan J Marugan, Liudmila Kulakova, Paul Shinn, Danielle van Leer, Wei Zheng, Osnat Herzberg","doi":"10.2174/1875397301206010093","DOIUrl":"https://doi.org/10.2174/1875397301206010093","url":null,"abstract":"The human pathogen Giardia lamblia is an anaerobic protozoan parasite that causes giardiasis, one of the most common diarrheal diseases worldwide. Although several drugs are available for the treatment of giardisis, resistance to these drugs has been reported and is likely to increase. The Giardia carbamate kinase (glCK) plays an essential role in Giardia metabolism and has no homologs in humans, making it an attractive candidate for anti-Giardia drug development. We have developed a luminescent enzyme coupled assay to measure the activity of glCK by quantitating the amount of ATP produced by the enzyme. This assay is homogeneous and has been miniaturized into a 1536-well plate format. A pilot screen against 4,096 known compounds using this assay yielded a signal-to-basal ratio of 11.5 fold and Z' factor of 0.8 with a hit rate of 0.9 % of inhibitors of glCK. Therefore, this Giardia lamblia carbamate kinase assay is useful for high throughput screening of large compound collection for identification of the inhibitors for drug development.","PeriodicalId":88232,"journal":{"name":"Current chemical genomics","volume":"6 ","pages":"93-102"},"PeriodicalIF":0.0,"publicationDate":"2012-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/af/ff/TOCHGENJ-6-93.PMC3565245.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31231196","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 18

Galectin-1 and Galectin-3 Mediate Protocadherin-24-Dependent Membrane Localization of β-catenin in Colon Cancer Cell Line HCT116. 半乳糖凝集素-1和半乳糖凝集素-3介导结肠癌细胞系HCT116中β-连环蛋白依赖原钙粘蛋白-24的膜定位。

Current chemical genomics Pub Date : 2012-01-01 Epub Date: 2012-09-20 DOI: 10.2174/1875397301206010018

Rui Ose, Osamu Oharaa, Takahiro Nagase

{"title":"Galectin-1 and Galectin-3 Mediate Protocadherin-24-Dependent Membrane Localization of β-catenin in Colon Cancer Cell Line HCT116.","authors":"Rui Ose, Osamu Oharaa, Takahiro Nagase","doi":"10.2174/1875397301206010018","DOIUrl":"https://doi.org/10.2174/1875397301206010018","url":null,"abstract":"Protocadherin-24 (PCDH24) is linked to the suppression of tumor growth and the inhibition of cell proliferation in the colon cancer cell line HCT116. We previously observed that β-catenin is localized to the plasma membrane when PCDH24 is expressed in these cells, but the molecular mechanisms by which PCDH24 induces the membrane localization of β-catenin remain largely unknown. To clarify these mechanisms, we identified molecules that interact with ectopically expressed PCDH24 in HCT116 cells using a HaloTag® pull-down assay. We found that galectin-1 and galectin-3 physically interact with PCDH24 and are retained at the plasma membrane in association with PCDH24 expression. A luciferase-based pull-down assay using HaloTag-fused galectins revealed that an intracellular region of PCDH24 (amino acids 1186-1280) is essential for this interaction. Furthermore, the over-expression of galectin-1 or -3, or the depletion of endogenous galectins by small interfering RNA modulates β-catenin translocation. We also revealed that the retention of galectin-1 and -3 at the plasma membrane results in the inactivation of PI3K activity. From these findings, we propose a model in which the galectin-anchoring activity of PCDH24 leads to the suppression of β-catenin signaling by the localization of β-catenin at the plasma membrane in PCDH24-expressing HCT116 colon cancer cells.","PeriodicalId":88232,"journal":{"name":"Current chemical genomics","volume":"6 ","pages":"18-26"},"PeriodicalIF":0.0,"publicationDate":"2012-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/65/1c/TOCHGENJ-6-18.PMC3480823.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31018185","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 28

Utilizing HaloTag Technology to Track the Fate of PCSK9 from Intracellular vs. Extracellular Sources. 利用HaloTag技术追踪细胞内与细胞外来源PCSK9的命运。

Current chemical genomics Pub Date : 2012-01-01 Epub Date: 2012-09-20 DOI: 10.2174/1875397301206010038

Xi Ai, Paul Fischer, Oksana C Palyha, Douglas Wisniewski, Brian Hubbard, Karen Akinsanya, Alison M Strack, Anka G Ehrhardt

{"title":"Utilizing HaloTag Technology to Track the Fate of PCSK9 from Intracellular vs. Extracellular Sources.","authors":"Xi Ai, Paul Fischer, Oksana C Palyha, Douglas Wisniewski, Brian Hubbard, Karen Akinsanya, Alison M Strack, Anka G Ehrhardt","doi":"10.2174/1875397301206010038","DOIUrl":"https://doi.org/10.2174/1875397301206010038","url":null,"abstract":"The function of a particular protein is dependent upon its localization and milieu. The ability to track the \"fate\" of a protein is a valuable tool to elucidate its function. We present the use of HaloTag technology to study the localization and fate of human Proprotein Convertase Subtilisin-like Kexin type 9 (PCSK9).The role of PCSK9 in the regulation of circulating low density lipoprotein-cholesterol (LDL-c) levels is ascribed to binding of circulating PCSK9 to the LDL receptor (LDLR) and subsequent lysosomal degradation of LDLR. However, hints in the literature indicate that intracellular PCSK9 may act on the LDLR, possibly during processing of newly synthesized protein. To address this question, the source and fate of intracellular PCSK9 requires further investigation.We applied HaloTag technology to distinguish the source of intracellular PCSK9 and showed that newly synthesized intracellular PCSK9 has unique localization from the PCSK9 after re-uptake. This suggests different functions of PCSK9 while interacting with the LDLR.","PeriodicalId":88232,"journal":{"name":"Current chemical genomics","volume":"6 ","pages":"38-47"},"PeriodicalIF":0.0,"publicationDate":"2012-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/6c/50/TOCHGENJ-6-38.PMC3480691.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31018187","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 7

A comparison of the performance and application differences between manual and automated patch-clamp techniques. 手动和自动膜片钳技术的性能和应用差异的比较。

Current chemical genomics Pub Date : 2012-01-01 Epub Date: 2012-12-28 DOI: 10.2174/1875397301206010087

Xiao Yajuan, Liang Xin, Li Zhiyuan

{"title":"A comparison of the performance and application differences between manual and automated patch-clamp techniques.","authors":"Xiao Yajuan, Liang Xin, Li Zhiyuan","doi":"10.2174/1875397301206010087","DOIUrl":"https://doi.org/10.2174/1875397301206010087","url":null,"abstract":"The patch clamp technique is commonly used in electrophysiological experiments and offers direct insight into ion channel properties through the characterization of ion channel activity. This technique can be used to elucidate the interaction between a drug and a specific ion channel at different conformational states to understand the ion channel modulators' mechanisms. The patch clamp technique is regarded as a gold standard for ion channel research; however, it suffers from low throughput and high personnel costs. In the last decade, the development of several automated electrophysiology platforms has greatly increased the screen throughput of whole cell electrophysiological recordings. New advancements in the automated patch clamp systems have aimed to provide high data quality, high content, and high throughput. However, due to the limitations noted above, automated patch clamp systems are not capable of replacing manual patch clamp systems in ion channel research. While automated patch clamp systems are useful for screening large amounts of compounds in cell lines that stably express high levels of ion channels, the manual patch clamp technique is still necessary for studying ion channel properties in some research areas and for specific cell types, including primary cells that have mixed cell types and differentiated cells that derive from induced pluripotent stem cells (iPSCs) or embryonic stem cells (ESCs). Therefore, further improvements in flexibility with regard to cell types and data quality will broaden the applications of the automated patch clamp systems in both academia and industry.","PeriodicalId":88232,"journal":{"name":"Current chemical genomics","volume":"6 ","pages":"87-92"},"PeriodicalIF":0.0,"publicationDate":"2012-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/11/7e/TOCHGENJ-6-87.PMC3549544.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31183574","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 49

Methods for Activity Analysis of the Proteins that Regulate Histone Methylation. 调节组蛋白甲基化的蛋白活性分析方法。

Current chemical genomics Pub Date : 2011-01-01 Epub Date: 2011-08-22 DOI: 10.2174/1875397301005010095

Amy M Quinn, Anton Simeonov

{"title":"Methods for Activity Analysis of the Proteins that Regulate Histone Methylation.","authors":"Amy M Quinn, Anton Simeonov","doi":"10.2174/1875397301005010095","DOIUrl":"https://doi.org/10.2174/1875397301005010095","url":null,"abstract":"The enzymes that regulate histone methylation states and the protein domains that recognize methylated histone residues have been implicated in a number of human diseases, including cancer, as a result of their ability to affect transcriptional changes by altering chromatin structure. These proteins are recognized as potential therapeutic targets for the treatment of diseases associated with epigenetic disruption; however, few inhibitors of their activity have been identified. The majority of histone demethylase and methyltransferase enzyme inhibitors have been discovered on the basis of their structural similarity to substrates or known inhibitors of enzymes with analogous mechanisms. The general lack of potency and specificity of these compounds indicates that novel chemotypes are needed to address the large number of recently discovered histone-modifying enzymes. High-throughput screening (HTS) allows rapid testing of chemically diverse small molecule libraries, provided assays amenable to HTS exist. Here we review the biochemical and cellular assays available for testing the proteins and enzymes that regulate histone methylation. Progress in the development of high-throughput, sensitive, and robust assays will enable discovery of small molecules for epigenetic therapy.","PeriodicalId":88232,"journal":{"name":"Current chemical genomics","volume":"5 Suppl 1","pages":"95-105"},"PeriodicalIF":0.0,"publicationDate":"2011-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/8c/02/TOCHGENJ-5-95.PMC3180180.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"30180365","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 14

Development and Implementation of a High Throughput Screen for the Human Sperm-Specific Isoform of Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDHS). 甘油醛3-磷酸脱氢酶(GAPDHS)人类精子特异性亚型高通量筛选方法的开发与实现

Current chemical genomics Pub Date : 2011-01-01 Epub Date: 2011-07-04 DOI: 10.2174/1875397301105010030

Jonathan Z Sexton, Polina V Danshina, David R Lamson, Mark Hughes, Alan J House, Li-An Yeh, Deborah A O'Brien, Kevin P Williams

{"title":"Development and Implementation of a High Throughput Screen for the Human Sperm-Specific Isoform of Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDHS).","authors":"Jonathan Z Sexton, Polina V Danshina, David R Lamson, Mark Hughes, Alan J House, Li-An Yeh, Deborah A O'Brien, Kevin P Williams","doi":"10.2174/1875397301105010030","DOIUrl":"https://doi.org/10.2174/1875397301105010030","url":null,"abstract":"Glycolytic isozymes that are restricted to the male germline are potential targets for the development of reversible, non-hormonal male contraceptives. GAPDHS, the sperm-specific isoform of glyceraldehyde-3-phosphate dehydrogenase, is an essential enzyme for glycolysis making it an attractive target for rational drug design. Toward this goal, we have optimized and validated a high-throughput spectrophotometric assay for GAPDHS in 384-well format. The assay was stable over time and tolerant to DMSO. Whole plate validation experiments yielded Z' values >0.8 indicating a robust assay for HTS. Two compounds were identified and confirmed from a test screen of the Prestwick collection. This assay was used to screen a diverse chemical library and identified fourteen small molecules that modulated the activity of recombinant purified GAPDHS with confirmed IC50 values ranging from 1.8 to 42 µM. These compounds may provide useful scaffolds as molecular tools to probe the role of GAPDHS in sperm motility and long term to develop potent and selective GAPDHS inhibitors leading to novel contraceptive agents.","PeriodicalId":88232,"journal":{"name":"Current chemical genomics","volume":"5 ","pages":"30-41"},"PeriodicalIF":0.0,"publicationDate":"2011-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/89/b8/TOCHGENJ-5-30.PMC3134944.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"30008332","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 11

Targets in epigenetics: inhibiting the methyl writers of the histone code. 表观遗传学的目标：抑制组蛋白密码的甲基写入。

Current chemical genomics Pub Date : 2011-01-01 Epub Date: 2011-08-22 DOI: 10.2174/1875397301005010072

Julianne M Yost, Ilia Korboukh, Feng Liu, Cen Gao, Jian Jin

引用次数: 0

Two High Throughput Screen Assays for Measurement of TNF-α in THP-1 Cells. 两种检测THP-1细胞中TNF-α的高通量筛选方法。

Current chemical genomics Pub Date : 2011-01-01 Epub Date: 2011-05-10 DOI: 10.2174/1875397301105010021

Kristin P Leister, Ruili Huang, Bonnie L Goodwin, Andrew Chen, Christopher P Austin, Menghang Xia

{"title":"Two High Throughput Screen Assays for Measurement of TNF-α in THP-1 Cells.","authors":"Kristin P Leister, Ruili Huang, Bonnie L Goodwin, Andrew Chen, Christopher P Austin, Menghang Xia","doi":"10.2174/1875397301105010021","DOIUrl":"https://doi.org/10.2174/1875397301105010021","url":null,"abstract":"Tumor Necrosis Factor-α (TNF-α), a secreted cytokine, plays an important role in inflammatory diseases and immune disorders, and is a potential target for drug development. The traditional assays for detecting TNF-α, enzyme linked immunosorbent assay (ELISA) and radioimmunoassay, are not suitable for the large size compound screens. Both assays suffer from a complicated protocol, multiple plate wash steps and/or excessive radioactive waste. A simple and quick measurement of TNF-α production in a cell based assay is needed for high throughput screening to identify the lead compounds from the compound library. We have developed and optimized two homogeneous TNF-α assays using the HTRF (homogeneous time resolved fluorescence) and AlphaLISA assay formats. We have validated the HTRF based TNF-α assay in a 1536-well plate format by screening a library of 1280 pharmacologically active compounds. The active compounds identified from the screen were confirmed in the AlphaLISA TNF-α assay using a bead-based technology. These compounds were also confirmed in a traditional ELISA assay. From this study, several beta adrenergic agonists have been identified as TNF-α inhibitors. We also identified several novel inhibitors of TNF-α, such as BTO-1, CCG-2046, ellipticine, and PD 169316. The results demonstrated that both homogeneous TNF-α assays are robust and suitable for high throughput screening.","PeriodicalId":88232,"journal":{"name":"Current chemical genomics","volume":"5 ","pages":"21-9"},"PeriodicalIF":0.0,"publicationDate":"2011-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/62/17/TOCHGENJ-5-21.PMC3106354.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"29915745","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 26

Chemical biology of lysine demethylases. 赖氨酸去甲基化酶的化学生物学。

Current chemical genomics Pub Date : 2011-01-01 Epub Date: 2011-08-22 DOI: 10.2174/1875397301005010062

Tom D Heightman

{"title":"Chemical biology of lysine demethylases.","authors":"Tom D Heightman","doi":"10.2174/1875397301005010062","DOIUrl":"https://doi.org/10.2174/1875397301005010062","url":null,"abstract":"Abnormal levels of DNA methylation and/or histone modifications are observed in patients with a wide variety of chronic diseases. Methylation of lysines within histone tails is a key modification that contributes to increased gene expression or repression depending on the specific residue and degree of methylation, which is in turn controlled by the interplay of lysine methyl transferases and demethylases. Drugs that target these and other enzymes controlling chromatin modifications can modulate the expression of clusters of genes, potentially offering higher therapeutic efficacy than classical agents acting on downstream biochemical pathways that are susceptible to degeneracy. Lysine demethylases, first discovered in 2004, are the subject of increasing interest as therapeutic targets. This review provides an overview of recent findings implicating lysine demethylases in a range of therapeutic areas including oncology, immunoinflammation, metabolic disorders, neuroscience, virology and regenerative medicine, together with a summary of recent advances in structural biology and small molecule inhibitor discovery, supporting the tractability of the protein family for the development of selective druglike inhibitors.","PeriodicalId":88232,"journal":{"name":"Current chemical genomics","volume":"5 Suppl 1","pages":"62-71"},"PeriodicalIF":0.0,"publicationDate":"2011-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/0f/86/TOCHGENJ-5-62.PMC3178875.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"30180362","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 23

Quantitative proteomic approaches to studying histone modifications. 定量蛋白质组学方法研究组蛋白修饰。

Current chemical genomics Pub Date : 2011-01-01 Epub Date: 2011-08-22 DOI: 10.2174/1875397301005010106

Barry M Zee, Nicolas L Young, Benjamin A Garcia

引用次数: 21