Synthesis and conformational analysis of an O-phosphotyrosine-containing α-helical peptide

Wade, John D.; Perich, John W.; McLeish, Michael J.; Ötvös, László; Tregear, Geoffrey W.

doi:10.1007/bf00128500

Cited by 7 publications

(3 citation statements)

References 21 publications

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“…LYS(59-81) was available from a previous study (McLeish et al, 1994). LYS(11-36), EG-LRLKIYKDTEGYYTIGIGHLLTKS, was assembled by Fmoc-solid phase synthesis on a MilliGen 9050 instrument using previously described methodology (Wade et al, 1995). It was cleaved from the solid support and simultaneously deprotected by treatment for 3.5 h with 95% trifluoroacetic acid/2.5% phenol/2.5% ethanethiol.…”

Section: Methodsmentioning

confidence: 99%

Conformational Analysis of LYS(11−36), a Peptide Derived from the β-Sheet Region of T4 Lysozyme, in TFE and SDS

et al. 1997

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The solution conformation of a peptide LYS(11-36), which corresponds to the beta-sheet region in T4 lysozyme, has been examined in aqueous solution, TFE, and SDS micelles by CD and 1H NMR spectroscopy. Secondary structure predictions suggest some beta-sheet and turn character in aqueous solution but predict a helical conformation in a more hydrophobic environment. The predictions were supported by the CD and NMR studies which showed the peptide to be relatively unstructured in aqueous solution, although there was some evidence of a beta-turn conformer which was maintained in 200 mM SDS and, to a lesser extent, in 50% TFE. The peptide was significantly helical in the presence of either 50% TFE or 200 mM SDS. TFE and SDS titrations showed that the peptide could form helical, sheet, or extended structure depending on the TFE or SDS concentration. The studies indicate that peptide environment is the determining factor in secondary structure adopted by LYS(11-36).

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Section: Methodsmentioning

confidence: 99%

Conformational Analysis of LYS(11−36), a Peptide Derived from the β-Sheet Region of T4 Lysozyme, in TFE and SDS

et al. 1997

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“…Synthesis of the biotinylated anti‐APCp2 (Fig. 1) was achieved using continuous flow Fmoc‐methodology essentially as described previously (14,15). The solid support was Fmoc‐PAL PEG‐PS (PerSeptive Biosystems, USA), and HBTU‐activated Fmoc‐amino acids were used throughout.…”

Section: Methodsmentioning

confidence: 99%

Structural and biosensor analyses of a synthetic biotinylated peptide probe for the isolation of adenomatous polyposis coli tumor suppressor protein complexes

Wade

Catimel

Faux

et al. 2001

The Journal of Peptide Research

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Large numbers of colon tumors stem from mutations in the gene coding for the production of the adenomatous polyposis coli (APC) tumor suppressor protein. This protein contains a coiled-coil N-terminal domain that is known to be responsible for homodimerization. Previous work by others has led to the design of a specific 54-residue anti-APC peptide (anti-APCp1) that dimerizes preferentially with this domain. We have undertaken the chemical synthesis of a modified form of this peptide (anti-APCp2) that bears a biotin moiety at its N-terminus for use in subsequent ligand-binding analysis studies. The peptide was subjected to comprehensive chemical characterization to confirm its purity. Secondary structural analysis by circular dichroism spectroscopy and Fourier transform infrared spectroscopy indicated that the peptide could assume a wide range of potential conformations, depending upon the precise microenvironment. Significantly, a stable alpha-helical structure was generated when the solvent conditions supported intramolecular salt-bridge formation along the helix barrel. The biotinylated anti-APCp2 was immobilized onto a streptavidin sensor surface, in a specific orientation leaving all amino acids available to form a coiled structure. In one experiment, injection of colonic cell lysate extracts (LIM1215) onto a size-exclusion column resulted in the isolation of a high molecular mass protein peak (> 600 kDa) that reacted specifically with the immobilized anti-APCp2 on the biosensor surface. In another experiment, a high molecular mass protein (M(r) > 250 kDa on SDS-PAGE) could be specifically immunoprecipitated from this peak using either the anti-APCp2 peptide or an anti-APC polyclonal antibody. This demonstrates the specific interaction between the anti-APCp2 peptide and native APC and highlights the potential use of the former peptide in a multidimensional micropreparative chromatographic/biosensor/proteomic protocol for the purification of APC alone and APC complexed with different biopolymers in various cell lines, and stages of tumor development.

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“…Several phosphopeptides (e.g. ETGTKAELLAKpYEATHK) have been synthesized with Boc-Tyr(PO 3 Bzl 2 )-OH through solid-phase methodology [13]. An O-phosphorylated tyrosine analog of human angiotensin II was chemically synthesized by using Fmoc-O-dimethylphosphono-L-tyrosine in a continuous-flow solid-phase synthesis.…”

Section: Phosphotyrosine Buildings Blocks With Two Phosphate Protectimentioning

confidence: 99%

Phosphopeptides - Chemical Synthesis, Analysis, Outlook and Limitations.

Tóth¹,

Kele²,

Váradi³

2007

COC

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Protein phosphorylation is one of the most important post-translational events in cell regulation and signal transduction. Since the isolation of phosphorylated peptides from biological sources is usually not feasible, there is a need for efficient chemical phosphorylation methods to allow the study of the role of phosphorylation/dephosphorylation in biological processes. The recent developments in phosphopeptide chemistry (special protecting groups for the phosphate moiety and new amidite reagents) have provided peptide chemists with a wide variety of different strategies applicable for work with the sensitive phosphoserine, phosphothreonine and phosphotyrosine-containing peptides. This has made possible the efficient chemical preparation of longer, multiply phosphorylated, possibly cell-permeable or fluorescent residue-bearing peptides.

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Synthesis and conformational analysis of an O-phosphotyrosine-containing α-helical peptide

Cited by 7 publications

References 21 publications

Conformational Analysis of LYS(11−36), a Peptide Derived from the β-Sheet Region of T4 Lysozyme, in TFE and SDS

Conformational Analysis of LYS(11−36), a Peptide Derived from the β-Sheet Region of T4 Lysozyme, in TFE and SDS

Structural and biosensor analyses of a synthetic biotinylated peptide probe for the isolation of adenomatous polyposis coli tumor suppressor protein complexes

Phosphopeptides - Chemical Synthesis, Analysis, Outlook and Limitations.

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