Capture of a Dimeric Intermediate during Transthyretin Amyloid Formation

Point mutations in the human plasma protein transthyretin are associated with the neurological disorder familial amyloidosis with polyneuropathy type 1. The disease is characterized by amyloid fibril deposits causing damage at the site of deposition. Substitution of two amino acids in the hydrophobi...

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Bibliographic Details
Published in:The Journal of biological chemistry 2001-10, Vol.276 (43), p.39592-39599
Main Authors: Olofsson, Anders, Ippel, Hans J., Baranov, Vladimir, Hörstedt, Per, Wijmenga, Sybren, Lundgren, Erik
Format: Article
Language:English
Subjects:
Amyloid - biosynthesis
Amyloid - ultrastructure
Amyloid Neuropathies, Familial - etiology
Anilino Naphthalenesulfonates
Asparagine - genetics
Dimerization
Fluorescent Dyes
Glutamic Acid - genetics
Guanidine - pharmacology
Humans
Models, Molecular
Mutation
Prealbumin - genetics
Prealbumin - metabolism
Protein Binding
Protein Denaturation
Protein Folding
Protein Structure, Secondary
Recombinant Proteins - metabolism
Staining and Labeling
Temperature
Thyroxine - metabolism
transthyretin
Valine - genetics
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Summary:Point mutations in the human plasma protein transthyretin are associated with the neurological disorder familial amyloidosis with polyneuropathy type 1. The disease is characterized by amyloid fibril deposits causing damage at the site of deposition. Substitution of two amino acids in the hydrophobic core of transthyretin lead to a mutant that was very prone to form amyloid. In addition, this mutant has also been shown to induce a toxic response on a neuroblastoma cell line. Renaturation of the transthyretin mutant at low temperature facilitated the isolation of an amyloid-forming intermediate state having the apparent size of a dimer. Increasing the temperature effectively enhanced the rate of interconversion from a partly denatured protein to mature amyloid. Using circular dichroism the β-sheet content of the formed mature fibrils was significantly lower than that of the native fold of transthyretin. Morphology studies using electron microscopy also indicated a temperature-dependent transformation from amorphous aggregates toward mature amyloid fibrils. In addition, 1-anilino-8-naphtalenesulfonate fluorescence studies suggested the loss of the thyroxin-binding channel within both the isolated intermediate and the mature fibrils.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M103599200