{"title":"2D‐DIGE analysis revealed reduced cytokeratin signaling in placenta with preeclampsia","authors":"R. Hass, M. Kirchner, B. Hollwitz, A. Scharf","doi":"10.1002/SITA.200500072","DOIUrl":"https://doi.org/10.1002/SITA.200500072","url":null,"abstract":"Preeclampsia represents a severe pregnancy disorder associated with premature delivery and fetal growth retardation which also involves certain imbalances of the placental maturation and the placental integration into the surrounding tissues. To characterize possible differences in the development of normal and preeclamptic placentae, two-dimensional SDS-PAGE of 10 normal placental protein homogenates was performed and compared to the protein pattern of homogenates from 10 preeclamptic placentae. Thus, acidic proteins particularly of about 56 kDa were identified in normal placentae which were nearly undetectable in protein homogenates of preeclamptic placentae. Peptide mass finger printing by MALDI identified cytokeratins, especially cytokeratin-10 as one of the differentially expressed protein. Quantitative data were obtained by two-dimensional difference-gel electrophoresis (2D DIGE). Labeling of normal and preeclamptic placental proteins with the fluorophors Cy5 and Cy3, respectively, and subsequent separation of these mixed samples by 2D DIGE revealed a differentially expressed protein spot at a 12.8-fold higher fluorescence intensity in normal placentae as compared to preeclamptic placentae. MALDI analysis of this differentially expressed protein spot identified cytokeratin. In order to verify these results in individual placenta probes, cytokeratin-10 Western blots were performed in 3 normal and preeclamptic placental homogenates of different gestational ages, respectively. Whereas a marked 56 kDa cytokeratin-10 expression appeared in all normal placentae, there was only little if any detectable cytokeratin-10 present in the preeclamptic placentae, respectively. These findings suggest that preeclampsia is accompanied by a significantly reduced cytokeratin signaling provided by 2D-DIGE-coupled MALDI analysis which represents a suitable technique to identify e. g. disease-related alterations in protein patterns.","PeriodicalId":88702,"journal":{"name":"Signal transduction","volume":"76 2","pages":"190-197"},"PeriodicalIF":0.0,"publicationDate":"2006-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/SITA.200500072","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50950412","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}
C. Ehrhardt, M. Schmolke, Andreas T. Matzke, Alexander Knoblauch, C. Will, V. Wixler, S. Ludwig
{"title":"Polyethylenimine, a cost-effective transfection reagent","authors":"C. Ehrhardt, M. Schmolke, Andreas T. Matzke, Alexander Knoblauch, C. Will, V. Wixler, S. Ludwig","doi":"10.1002/SITA.200500073","DOIUrl":"https://doi.org/10.1002/SITA.200500073","url":null,"abstract":"In search for a cheap and effective transfection reagent we used the positively charged polyplex-forming compound polyethylenimine (PEI). This compound is commercially available from different companies either as a non-modified chemical reagent or with additives as a more cost intensive transfection reagent. Here we used the non-modified PEI reagent to optimize transfection protocols for different cell-lines. With these optimized conditions we were able to transiently transfect a number of cell-lines up to 40–90%.","PeriodicalId":88702,"journal":{"name":"Signal transduction","volume":"15 1","pages":"179-184"},"PeriodicalIF":0.0,"publicationDate":"2006-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/SITA.200500073","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50950420","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}
{"title":"Physical coupling between lipids and proteins: a paradigm for cellular control","authors":"O. Ces, X. Mulet","doi":"10.1002/SITA.200500079","DOIUrl":"https://doi.org/10.1002/SITA.200500079","url":null,"abstract":"The long held view of biological lipids acting as passive building blocks in biological membranes has now been superseded by the notion that the structure and composition of the membrane plays a crucial role in the regulation of protein function and numerous other biological processes. This may be the result of specific protein-lipid recognition phenomena or alternatively regulation by sensing properties of the membrane such as the stored curvature elastic stress with the latter being the focus of the review.","PeriodicalId":88702,"journal":{"name":"Signal transduction","volume":"26 5","pages":"112-132"},"PeriodicalIF":0.0,"publicationDate":"2006-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/SITA.200500079","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50950542","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}
{"title":"The role of phosphoinositides in mast cell signalling","authors":"Richard D. Byrne, B. Larijani","doi":"10.1002/SITA.200500074","DOIUrl":"https://doi.org/10.1002/SITA.200500074","url":null,"abstract":"The release of pre-formed mediators such as histamine from mast cells and basophils is an integral part of the normal immune response to infection by parasites. This exocytosis is also characteristic response in a number of disease states including asthma, which, due to their prevalence in western society are becoming of increasing clinical importance. In an effort to tackle this growing problem much work has gone into unlocking the mechanisms through which mast cells function in health and disease. To date we have learned a lot about the various proteins that regulate degranulation. However, our knowledge on the contribution of lipids to this process is less clear. This review will discuss the role of phospholipids, particularly the phosphoinositides (PIs) in the processes that regulate mast cell exocytosis.","PeriodicalId":88702,"journal":{"name":"Signal transduction","volume":"6 1","pages":"81-91"},"PeriodicalIF":0.0,"publicationDate":"2006-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/SITA.200500074","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50950429","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}
E. Tate, Paul W. Bowyer, K. Brown, Deborah F. Smith, A. Holder, R. Leatherbarrow
{"title":"Peptide‐based inhibitors of N‐myristoyl transferase generated from a lipid/combinatorial peptide chimera library","authors":"E. Tate, Paul W. Bowyer, K. Brown, Deborah F. Smith, A. Holder, R. Leatherbarrow","doi":"10.1002/SITA.200500084","DOIUrl":"https://doi.org/10.1002/SITA.200500084","url":null,"abstract":"Peptide aptamers are powerful chemical genetic tools for the dissection of biological networks, but their application to in vivo systems has been limited by the challenging problem of targeting peptides to a specific site on a single target protein. Here we present our initial research on a novel technique for targeting combinatorial peptide aptamers to a protein binding-site using a small-molecule binding-partner (or ‘Trojan horse’). Novel peptide-based inhibitors for Plasmodium falciparum myristoyl-CoA:protein N-myristoyl transferase (PfNMT) have been selected from a one-bead one-compound library using a high-throughput on-bead screening methodology, targeted to the active site of NMT with a myristate (C14 : 0 fatty acid) substrate analogue. From an initial screen of an unbiased 130321-compound library of lipid/combinatorial peptide chimeras, we have selected 6-mer peptides in an on-bead assay which show NMT inhibition with IC50 values ranging down to low micromolar.","PeriodicalId":88702,"journal":{"name":"Signal transduction","volume":"66 1","pages":"160-166"},"PeriodicalIF":0.0,"publicationDate":"2006-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/SITA.200500084","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50950664","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}
{"title":"Characterisation of lipids in cell signalling and membrane dynamics by nuclear magnetic resonance spectroscopy and mass spectrometry","authors":"Marie Garnier, E. Dufourc, B. Larijani","doi":"10.1002/SITA.200500077","DOIUrl":"https://doi.org/10.1002/SITA.200500077","url":null,"abstract":"Signalling lipids regulate cellular events directly by recruiting and/or activating specific proteins or indirectly by locally modifying the physical properties of the membrane. These events trigger the co-localisation and interaction of proteins on membranes. This in turn effects the function and the properties of the proteins. To enable a comprehensive study of the complex and spatially heterogeneous natural membrane, complementary analytical tools need to be applied. Spectroscopy tools such as nuclear magnetic resonance spectroscopy and mass spectrometry provide the sensitivity and the precision required for detecting low amounts of highly transient signalling lipids, but also determine their effect on membrane structure. Specific examples are used to illustrate the application of liquid NMR spectroscopy, solid state NMR spectroscopy and electrospray ionisation mass spectrometry. The main advantages and limitations of these tools are discussed in this review.","PeriodicalId":88702,"journal":{"name":"Signal transduction","volume":"101 1","pages":"133-143"},"PeriodicalIF":0.0,"publicationDate":"2006-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/SITA.200500077","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50950476","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}
{"title":"Nuclear signalling through phospholipase C and phosphatidyl 4,5-bisphosphate","authors":"K. Ho, D. Mann","doi":"10.1002/SITA.200500078","DOIUrl":"https://doi.org/10.1002/SITA.200500078","url":null,"abstract":"","PeriodicalId":88702,"journal":{"name":"Signal transduction","volume":"9 3","pages":"92-100"},"PeriodicalIF":0.0,"publicationDate":"2006-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/SITA.200500078","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50950530","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}
{"title":"Phospholipid signalling: mediators in need of interdisciplinary techniques","authors":"R. Woscholski","doi":"10.1002/SITA.200690018","DOIUrl":"https://doi.org/10.1002/SITA.200690018","url":null,"abstract":"","PeriodicalId":88702,"journal":{"name":"Signal transduction","volume":"52 2","pages":"77-79"},"PeriodicalIF":0.0,"publicationDate":"2006-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/SITA.200690018","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50951346","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}
{"title":"Interactions of synaptojanin","authors":"E. Rosivatz","doi":"10.1002/SITA.200500076","DOIUrl":"https://doi.org/10.1002/SITA.200500076","url":null,"abstract":"Synaptojanin is a phosphoinositide phosphatase that plays a crucial role in synaptic vesicle recycling. The focus of this review is on the major direct interacting partners of synaptojanin with respect to recognition motifs in synaptojanin isoforms and splice variants. Those proteins that bind synaptojanin are mainly involved in endocytosis and function at the interface between actin and the endocytic machinery. Some interaction partners underline synaptojanin's emerging role in signaling and pathophysiological processes.","PeriodicalId":88702,"journal":{"name":"Signal transduction","volume":"101 9","pages":"101-111"},"PeriodicalIF":0.0,"publicationDate":"2006-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/SITA.200500076","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50950468","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}