|A Sensitive Fluorimetric Assay for Detection of ß-Secretase Activity Using a Novel FRET Peptide Substrate|
Xudong Zhu, Xing Han, Manpreet Mann, Rich Meyer, Xiaohe Tong, Anita Hong and Vera Rakhmanova
In order to facilitate high throughput screening of AD drug candidates, we have developed a new SensoLyte™ 520 b-secretase assay kit using a fluorescence resonance energy transfer (FRET) peptide, HiLyte Fluor™ 488-Glu-Val-Asn-Leu-Asp-Ala-Glu-Phe-Lys(QXL™ 520)-OH. The longer excitation and emission wavelengths of HiLyte Fluor™488 minimize the interference from autofluorescence and absorbance of test compounds.
|Glycoprotein Labeling and Detection: Novel Click Chemistry-based Applications for gel Electrophoresis, Flow Cytometry and Fluorescence Microscopy|
Brian Agnew, Nancy Ahnert, Suzanne Buck, Scott Clarke, Courtenay Hart, Kapil Kumar, and Tamara Nyberg
We demonstrate highly-selective and sensitive labeling methods for the detection of specific glycoprotein subclasses, including cell surface N- and O-linked glycoproteins and intracellular O-GlcNAc modified proteins, utilizing the copper-catalyzed cycloaddition reaction between azides and alkynes, or click chemistry.
|PRIMe: Platform for RIKEN Metabolomics|
Atsushi Fukushima, Miyako Kusano, Kenji Akiyama, Takeshi Obayashi, Takayuki Tohge, Masami Yokota Hirai, Shigehiko Kanaya, Masanori Arita, Yoko Shinbo, Kazuo Shinozaki, Tetsuya Sakurai and Kazuki Saito
We have developed a web-based database, "PRIMe (Platform for RIKEN Metabolomics)," which contains powerful tools for researchers to analyze gene co-expression data and mass spectral data. PRIMe has been developed with the main aim of facilitating integrated analysis for transcriptomics and metabolomics.
|Proteomic Profiling in Defining Chemoresistant Breast Cancer|
Chuthapisith S, Layfield R, Kerr I, Hughes C and Eremin O
This study aims to identify protein profiles in breast cancer cells as predictors of chemoresistance by using two-dimensional gel electrophoresis and MALDI-TOF peptide mass fingerprinting. Our findings provide further insights into the complex mechanisms of chemoresistance, as well as representing an attractive starting point for the identification of potential protein biomarkers to predict response to chemotherapy in breast cancer in vivo.
|Kinetic Constants and Sample-to-Sample Variation in the Rate of Metabolism of two or more Substrates for Human Liver Microsomal CYP1A2, CYP2B6, CYP2C8, CYP2D6 and CYP3A4/5|
Zell Woodworth, L. Anne Dwyer, Lisa Collins, Terry Graves, Stephanie Helmstetter, Brian Ogilvie, Clayton Otwell, Chad Pope, Tiffin Ramsey, Phyllis Yerino and Andrew Parkinson
Marker substrates for certain CYPs have changed for several reasons, including the need for improved selectivity and sensitivity, reliable substrates for LC/MS/MS analysis, and substrates that provide greater repeatability. The objectives of this study were to compare sample-to-sample variation in cytochrome P450 enzymatic rates between two or more CYP-specific substrates and to determine Michaelis-Menten kinetic constants for these same reactions using a pool of human liver microsomes.
|Evaluation of Different Interpretation Strategies to Discover PTM in MS/MS Peptide Fragmentation Data|
Daniel Chamrad, Gerhard Korting, Ken Fantom, Andy West, Klaus Schneider, Ulrike Schweiger-Hufagel, Herbert Thiele and Martin Bluggel
The phenomenon of acquired high quality MS/MS spectra that can not be explained within typical sequence database searches is well known. Although protein identification was successful it is manually very laborious and in most cases even impossible to match these spectra with any suggested protein sequence. The procedure of second pass searches has been developed to overcome this problem. Here we report from our in house developed tool PTM-Explorer.
|Towards Understanding of Ryegrass-Endophyte Symbiosis through Data Integration of Transcriptome and Metabolome|
Mingshu Cao, Linda Johnson, Albert Koulman, Geoff Lane and Susanne Rasmussen
Combined with extensive bioinformatics analyses, our joint analysis of transcriptomics and metabolomics data has generated interesting findings which led to further laboratory verification, such as potential genes associated with alkaloid production.
|Herbal Metabolic Profiling of Raw and Steamed Panax notoginseng|
Eric C.Y. Chan, Swee-Lee Yap, Aik-Jiang Lau, Pay-Chin Leow, Ding-Fung Toh, and Hwee-Ling Koh
At present, metabolite profiling is of growing importance in herbal medicine fields such as breeding, formulation, quality control and clinical trials. Herbal metabolic profiling allows direct detection of down-stream derivatives of metabolites, arising from herbal formulation process, using metabolite profiling ultra performance liquid chromatography time-of-flight mass spectrometry.
|Metabolic Profiling of Human Blood Plasma by Combined Ultra Performance Liquid Chromatography / Mass Spectrometry|
Steve Bruce, Pär Jonsson, Henrik Antti, Olivier Cloarec, Stefan Marklund and Thomas Moritz
Metabolic profiling is concerned with the analysis of low molecular weight compounds present in complex samples such as plasma. Analytical techniques such as UPLC/MS are proving to be powerful tools for metabolic profiling on complex samples such as plasma. Here we describe a protocol of metabolic profiling analysis on plasma by UPLC/MS.