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TL;DR: The beta-elimination/Michael addition method was successfully used to identify phosphorylated sequences of ovalbumin and human Stat1 by in-gel derivatization with 2-phenylethanethiol and subsequent peptide mass fingerprint analysis of the trypsin digests.
Abstract: The identification of phosphorylation sites is essential for a full understanding of the cellular functions of proteins. However, mass spectrometric analysis is often hampered by the low abundance of phosphoproteins, the difficulty of obtaining full sequence coverage by specific proteolysis reactions, and the low ionization efficiency of phosphopeptides compared with their non-phosphorylated analogs. In the present work a beta-elimination/Michael addition was used to replace the phosphate groups of pSer or pThr by a group which gives rise to an enhanced ionization efficiency. In order to find optimum reaction conditions, beta-elimination/Michael addition was examined using phosphorylated model peptides. Whereas complete elimination of phosphate could be achieved by treatment with barium hydroxide in organic solvents such as ethanol or acetonitrile, the yield of the Michael adduct strongly depended on the nucleophile and the peptide sequence. Reaction with 2-phenylethanethiol, p-bromophenethylamine and ethylenediamine clearly resulted in products showing higher matrix-assisted laser desorption/ionization (MALDI) signal intensities compared with those of the corresponding phosphorylated precursors. The method was successfully used to identify phosphorylated sequences of ovalbumin and human Stat1 by in-gel derivatization with 2-phenylethanethiol and subsequent peptide mass fingerprint analysis of the trypsin digests. Copyright © 2004 John Wiley & Sons, Ltd.
50 citations
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18 Mar 2005TL;DR: In this article, a method for separating polynucleotides such as DNA, RNA and PNA from solutions containing polyn nucleotides by reversibly binding the polynuclotides to a solid surface, such as a magnetic microparticle, was proposed.
Abstract: The invention relates to methods of separating polynucleotides, such as DNA, RNA and PNA, from solutions containing polynucleotides by reversibly binding the polynucleotides to a solid surface, such as a magnetic microparticle.
50 citations
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16 May 2007TL;DR: In this paper, the authors present an embodiment of a system for analysis of an analyte, which includes an excitation light source configured to induce an evanescent wave excitation of a fluorescently labeled molecule near the access to the transparent substrate and a detector for detecting the fluorescently labelled molecule.
Abstract: An embodiment generally relates to a system for analysis of an analyte. The system can include a transparent substrate. The system also includes an excitation light source configured to induce an evanescent wave excitation of a fluorescently labeled molecule near the access to the transparent substrate and a detector for detecting the fluorescently labeled molecule.
50 citations
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TL;DR: This study has identified and characterised the expression profiles of members of the anhydrobiotic gene set in A. avenae and demonstrates the potential of RNAi for the analysis of anHydrobiosis and provides the first genetic data to underline the importance of effective antioxidant systems in metazoan desiccation tolerance.
Abstract: Background
Some organisms can survive extreme desiccation by entering a state of suspended animation known as anhydrobiosis The free-living mycophagous nematode Aphelenchus avenae can be induced to enter anhydrobiosis by pre-exposure to moderate reductions in relative humidity (RH) prior to extreme desiccation This preconditioning phase is thought to allow modification of the transcriptome by activation of genes required for desiccation tolerance
50 citations
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TL;DR: A formula is devised that incorporates both resolution and separation time along with a third factor, peak distribution, that provides for both ordinal and cardinal ranking of chromatograms and can be used to establish historical norms.
49 citations
Authors
Showing all 1521 results
Name | H-index | Papers | Citations |
---|---|---|---|
Richard A. Gibbs | 172 | 889 | 249708 |
Friedrich C. Luft | 113 | 1095 | 47619 |
Alexander N. Glazer | 71 | 208 | 21068 |
Vineet Bafna | 68 | 236 | 42574 |
Kevin R. Coombes | 63 | 308 | 23592 |
Darryl J. Pappin | 61 | 170 | 29409 |
Mark D. Johnson | 60 | 289 | 16103 |
György Marko-Varga | 56 | 409 | 12600 |
Paul Thomas | 56 | 128 | 44810 |
Gerald Zon | 55 | 256 | 11126 |
Michael W. Hunkapiller | 51 | 130 | 29756 |
Bjarni V. Halldorsson | 51 | 145 | 13180 |
David H. Hawke | 50 | 157 | 9824 |
Ellson Y. Chen | 50 | 71 | 28836 |
Sridhar Hannenhalli | 49 | 162 | 21959 |