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Monitoring fibril formation of the N‐terminal domain of PABPN1 carrying an alanine repeat by tryptophan fluorescence and real‐time NMR
Author(s) -
Rohrberg Julia,
Sachs Rolf,
Lodderstedt Grit,
Sackewitz Mirko,
Balbach Jochen,
Schwarz Elisabeth
Publication year - 2008
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2008.04.002
Subject(s) - fibril , kinetics , chemistry , fluorescence , alanine , biophysics , nuclear magnetic resonance spectroscopy , fluorescence spectroscopy , tryptophan , crystallography , amino acid , biochemistry , stereochemistry , biology , physics , quantum mechanics
Intranuclear fibrils due to poly‐alanine expansions in the N‐terminal domain of the poly(A) binding protein PABPN1 correlate with the disease oculopharyngeal muscular dystrophy (OPMD). For monitoring fibril formation by fluorescence and real‐time NMR spectroscopy, tryptophans were introduced either into the middle or C‐terminal of the poly‐alanine segment. The kinetics of fibril formation which were monitored by fluorescence spectroscopy were matched by real‐time NMR kinetics. Our results show that fibril formation is concomitant with the burial of the tryptophans in the fibrillar core. Since no soluble pre‐fibrillar intermediate(s) was detected, fibril formation of this domain may be regarded as a two state conversion from an unfolded soluble into folded insoluble species.