|Alignment of 1H-NMR data using a Generalized Fuzzy Hough Transform|
Erik Alm, Leonard Csenki, Ralf J.O. Torgrip, K. Magnus Åberg, Lars I. Nord, Ina Schuppe-Koistinen and Johan Lindberg
In metabolic profiling, multivariate data analysis techniques are used to interpret 1D 1H–NMR data. Multivariate data analysis techniques require that peaks are located in the same variables in every spectrum – this requirement is not met in native NMR spectra. Current state-of-the-art alignment algorithms are unable to align peaks when the spatial order of the peaks changes. We present the Fuzzy Generalized Hough Transform alignment which solves the alignment problem.
|Specificity of small molecule inhibitors for deubiquitinating enzymes in living cells assessed by activity-based proteomics|
*Mikael Altun, *Holger B. Kramer, *Lianne Willems, *Mukram Mackeen, *Edward Kogan, *Rebecca Konietzny, *Cynthia Wright, *Roman Fischer, #Benjamin Nicholson, *Benedikt M. Kessler
Small molecular compounds (PR-619 and P22077) was assessed for their abilities to inhibit DUB function in crude extracts and in cells. Activity-based profiling combined with quantitative mass spectrometry revealed the inhibitory capacity of a broad range of DUBs by the PR-619, whereas P22077 showed specificity towards subsets of DUBs including USP7 in cells. Our results demonstrate the usefulness of activity-based quantitative proteomics to monitor inhibition of endogenous DUBs in vivo.
|Capture Compounds towards a targeted reduction of proteome complexity|
Erik Duelsner, Aysel Alici, Christian Jurinke, Hubert Koester
Capture Compound Mass Spectrometry (CCMS) enables the enrichment of proteins based on their functionality. CCMS is commercialized as kits for research applications and in collaborations with pharmaceutical companies. The focus is on investigating mechanisms of drug action and avoidance of toxic effects of small molecule drugs.
|Quantitative phosphoproteome analysis for the detection of early alterations of signal transduction by dioxin (TCDD)|
Melanie Schulz, Ulrich Andrae, Martin R. Larsen
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is an extremely toxic pollutant and regarded the most potent chemical carcinogen in experimental animals. Most of its biological effects are mediated by binding of TCDD to the cytosolic Ah receptor (AHR). In addition, TCDD cause AHR-dependent alterations in signal transduction that are independent of changes in gene expression. In this study, we have therefore conducted a quantitative phospho proteomic study on TCDD-induced alterations.
|Sialylation and Metastasis: from tumor-associated antigens discovery to therapeutic development|
Giuseppe Palmisano, Rikke Leth-Larsen and Martin R. Larsen
Aberrant sialylation affects the metastatic behaviour of cancer cells.We have quantified the sialylated cell surface glycoproteins from cells with different metastatic behaviour.The membrane proteins from isogenic human cell lines were analyzed by a method based on TiO2-HILIC-MS.1200 sialylated glycosites, 116 enzymes involved in the glycoconjugate metabolism and for the first time 4 glycosylation sites of Her2 receptor were identified, showing the importance of this tool for discovery and thera
|Proteomic non-small cell lung carcinoma biomarker screening in bronchoalveolar lavage fluid|
Tonio Oumeraci 1, Bernd Schmidt 3, Thomas Wolf 1,4, Marc Zapatka 4, Andreas Pich 2, Benedikt Brors 4, Roland Eils 4,5, Brigitte Schlegelberger 1, Nils von Neuhoff 1
Using a standardized method to acquire MALDI-TOF proteome profile spectra of bronchoalveolar lavage fluid (BALF), we have shown the upregulation of histatin 3 and calgranulin C in a small pilot cohort of NSCLC patients. This pilot study serves to demonstrate that it is feasible to screen a larger NSCLC patient cohort for BALF proteome level biomarkers in a clinical setting.
|Some thiazole derivatives as potent antiproliferative agents with CDK2/Cycin E/A inhibitory activities|
Mahesh Chhabria1, Shailesh Patel*1, Maulik Suthar2
We report synthesis and biological evaluation of novel thiazole derivatives. High throughput screening of various compounds for their anti-proliferative activity led to the identification of thiazolo-thiones as potential candidate for Lung cancer (NCl-H23, NClH510A, NCI-H522) and breast cancer (MDA-MB-453/231/468, MCF-7). The most potent compounds were also screened for their inhibitory activity against CDK2/ Cyclin E/A which are considered as promising targets for lung and breast cancer.
|Predicting hepatotoxicity: Reactive metabolite trapping using glutathione and freshly isolated hepatocytes|
Birks, V., Webber, G., Geoffroy, S., Cole, R., and Wood, S.
This poster presents our results to date using clozapine (a compound known to be associated with GSH-adduct formation) as substrate and using stable isotope GSH (GSH13C2,15N) to enhance specificity. In addition, all analyses have been conducted using an Waters Acquity UPLC-MS/MS. Results we have obtained in hepatocytes are compared against findings using human liver microsomes (HLM).
|A simple, fast and quantitative single-step dead-cell indicator for flow cytometry|
Jixiang Liu, Jolene Bradford, Chris Langsdorf
We have evaluated a series of new compounds for dead cell stain and identified a new product, SYTOX® AADvancedTM dead cell stain, which demonstrates improved properties over 7-AAD. These properties make the SYTOX® AADvancedTM dead cell stain a simple, fast and quantitative single-step no-wash dead-cell indicator as well as ideal for use in multicolor application requiring DNA content.