Neighbored phosphorylation sites as PHF-tau specific markers in Alzheimer’s disease

Singer, David; Lehmann, Jörg; Hanisch, Katja; Härtig, Wolfgang; Hoffmann, Ralf

doi:10.1016/j.bbrc.2006.05.201

Cited by 31 publications

(53 citation statements)

References 36 publications

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“…The peptides were synthesized on solid phase and purified to homogeneity by RPLC, 20,35 with purity confirmed by CZE. 21 The peptides were first screened one at a time with the peptide Syntide 2 (St) of similar (but not overlapping) m = 1508 Da as the internal calibrant.…”

Section: Experimental Methodsmentioning

confidence: 99%

Ion Mobility Separation of Isomeric Phosphopeptides from a Protein with Variant Modification of Adjacent Residues

et al. 2011

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Ion mobility spectrometry (IMS), and particularly differential or field asymmetric waveform IMS (FAIMS), was recently shown capable of separating post-translationally modified peptides with variant PTM localization. However, that work was limited to a model peptide with Ser phosphorylation on fairly distant alternative sites. Here, we demonstrate that FAIMS (coupled to ESI/MS) can broadly baseline-resolve variant phosphopeptides from a biologically modified human protein, including those involving phosphorylation of different residues and adjacent sites that challenge existing MS/MS methods most. Singly and doubly phosphorylated variants can be resolved equally well and identified without dissociation, based on accurate separation properties. The spectra change little over a range of infusion solvent pH, hence the present approach should be viable in conjunction with chromatographic separations using mobile phase gradients.

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Section: Experimental Methodsmentioning

confidence: 99%

Ion Mobility Separation of Isomeric Phosphopeptides from a Protein with Variant Modification of Adjacent Residues

et al. 2011

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“…The concentrations of the purified peptides as well as the synthetic ones were determined using tyrosine absorption (e 280 = 1290 M -1 cm (Fields and Noble 1990) using a Syro2000 multiple peptide synthesizer (MultiSynTech GmbH, Witten, Germany) (Singer et al 2006). After completion of the peptide synthesis, the peptides were cleaved with 5% water, 4% m-cresol, 5% thioanisole and 2% ethanedithiol in trifluoroacetic acid (TFA) at room temperature for 4 h and precipitated with cold diethyl ether.…”

Section: Sequence Analysismentioning

confidence: 99%

ChBac3.4: A Novel Proline-Rich Antimicrobial Peptide from Goat Leukocytes

Shamova

Орлов

Stegemann

et al. 2009

Int J Pept Res Ther

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“…They were synthesized as C-terminal amides on a Syro 2000 multiple peptide synthesizer (MultiSynTech GmbH, Witten, Germany) equipped with a 48-vessel reaction block using standard 9-fluorenylmethoxycarbonyl(Fmoc)/tert.-butyl chemistry [21]. Sites to be phosphorylated were incorporated without side-chain protection as Fmoc-Ser-OH or Fmoc-Thr-OH and globally phosphorylated by the phosphoamidite approach [20,22]. After cleavage with TFA the crude peptides were purified on a Jupiter C 18 -column (Phenomenex, Torrance, USA) with a linear aqueous acetonitrile gradient in the presence of 0.1% v/v TFA.…”

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confidence: 99%

Separation of multiphosphorylated peptide isomers by CZE

Muetzelburg

Hoffmann

2008

Electrophoresis

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A separation of mono, doubly and triply phosphorylated isomers was developed with CZE with an aqueous electrolyte containing 3.9 mol/L formic acid and 30% v/v trifluoroethanol. Thus a mixture of ten phosphopeptides corresponding to the human tau sequence 226-240 was separated within 70 min. Although peptides with different phosphorylation degrees, i.e. 0-3 phosphate groups, were well separated, some of the phosphopeptide isomers containing one or two phosphate groups were only partially separated. The electrolyte system is compatible with both MALDI- and ESI-MS, allowing a direct coupling, and thus could have some interesting applications in proteomics.

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Neighbored phosphorylation sites as PHF-tau specific markers in Alzheimer’s disease

Cited by 31 publications

References 36 publications

Ion Mobility Separation of Isomeric Phosphopeptides from a Protein with Variant Modification of Adjacent Residues

Ion Mobility Separation of Isomeric Phosphopeptides from a Protein with Variant Modification of Adjacent Residues

ChBac3.4: A Novel Proline-Rich Antimicrobial Peptide from Goat Leukocytes

Separation of multiphosphorylated peptide isomers by CZE

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