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@ARTICLE{Dammers:276619,
      author       = {Dammers, Christina and Gremer, Lothar and Neudecker,
                      Philipp and Demuth, Hans-Ulrich and Schwarten, Melanie and
                      Willbold, Dieter},
      title        = {{P}urification and {C}haracterization of {R}ecombinant
                      {N}-{T}erminally {P}yroglutamate-{M}odified {A}myloid-β
                      {V}ariants and {S}tructural {A}nalysis by {S}olution {NMR}
                      {S}pectroscopy},
      journal      = {PLoS one},
      volume       = {10},
      number       = {10},
      issn         = {1932-6203},
      address      = {Lawrence, Kan.},
      publisher    = {PLoS},
      reportid     = {FZJ-2015-06964},
      pages        = {e0139710},
      year         = {2015},
      abstract     = {Alzheimer’s disease (AD) is the leading cause of dementia
                      in the elderly and is characterized by memory loss and
                      cognitive decline. Pathological hallmark of AD brains are
                      intracellular neurofibrillary tangles and extracellular
                      amyloid plaques. The major component of these plaques is the
                      highly heterogeneous amyloid-β (Aβ) peptide, varying in
                      length and modification. In recent years
                      pyroglutamate-modified amyloid-β (pEAβ) peptides have
                      increasingly moved into the focus since they have been
                      described to be the predominant species of all N-terminally
                      truncated Aβ. Compared to unmodified Aβ, pEAβ is known to
                      show increased hydrophobicity, higher toxicity, faster
                      aggregation and β-sheet stabilization and is more resistant
                      to degradation. Nuclear magnetic resonance (NMR)
                      spectroscopy is a particularly powerful method to
                      investigate the conformations of pEAβ isoforms in solution
                      and to study peptide/ligand interactions for drug
                      development. However, biophysical characterization of pEAβ
                      and comparison to its non-modified variant has so far been
                      seriously hampered by the lack of highly pure recombinant
                      and isotope-enriched protein. Here we present, to our
                      knowledge, for the first time a reproducible protocol for
                      the production of pEAβ from a recombinant precursor
                      expressed in E. coli in natural isotope abundance as well as
                      in uniformly [U-15N]- or [U-13C, 15N]-labeled form, with
                      yields of up to 15 mg/l E. coli culture broth. The chemical
                      state of the purified protein was evaluated by RP-HPLC and
                      formation of pyroglutamate was verified by mass
                      spectroscopy. The recombinant pyroglutamate-modified Aβ
                      peptides showed characteristic sigmoidal aggregation
                      kinetics as monitored by thioflavin-T assays. The quality
                      and quantity of produced pEAβ40 and pEAβ42 allowed us to
                      perform heteronuclear multidimensional NMR spectroscopy in
                      solution and to sequence-specifically assign the backbone
                      resonances under near-physiological conditions. Our results
                      suggest that the presented method will be useful in
                      obtaining cost-effective high-quality recombinant pEAβ40
                      and pEAβ42 for further physiological and biochemical
                      studies.},
      cin          = {ICS-6},
      ddc          = {500},
      cid          = {I:(DE-Juel1)ICS-6-20110106},
      pnm          = {553 - Physical Basis of Diseases (POF3-553)},
      pid          = {G:(DE-HGF)POF3-553},
      typ          = {PUB:(DE-HGF)16},
      UT           = {WOS:000362499200055},
      pubmed       = {pmid:26436664},
      doi          = {10.1371/journal.pone.0139710},
      url          = {https://juser.fz-juelich.de/record/276619},
}