Abstract
We describe the expression and purification of a model amyloidogenic peptide comprising residues 105-115 of human transthyretin (TTR105-115). Recombinant TTR105-115, which does not contain any non-native residues, was prepared as part of a fusion protein construct with a highly soluble B1 immunoglobulin binding domain of protein G (GB1), with typical yields of similar to 4 mg/L of uniformly C-13,N-15-enriched HPLC-purified peptide per liter of minimal media culture. Amyloid fibrils formed by recombinant TTR105-115 were characterized by transmission electron microscopy and solid-state NMR spectroscopy, and found to be comparable to synthetic TTR105-115 fibrils. These results establish recombinant TTR105-115 as a valuable model system for the development of new solid-state NMR techniques for the atomic-level characterization of amyloid architecture. (C) 2009 Elsevier Inc. All rights reserved.
Original language | English |
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Pages (from-to) | 101-108 |
Number of pages | 8 |
Journal | Protein Expression and Purification |
Volume | 70 |
Issue number | 1 |
DOIs | |
Publication status | Published - Mar 2010 |
Keywords / Materials (for Non-textual outputs)
- Fusion protein
- GB1
- Recombinant human transthyretin 105-115
- Escherichia coli BL21(DE3)
- Ni2+ affinity chromatography
- Amyloid fibrils
- Solid-state NMR spectroscopy
- ALPHA-SYNUCLEIN FIBRILS
- BETA-SHEET STRUCTURE
- HUMAN PRION PROTEIN
- ANGLE-SPINNING NMR
- ESCHERICHIA-COLI
- HIGH-RESOLUTION
- MOLECULAR-CONFORMATION
- STRUCTURAL MODEL
- RESIDUES
- SYSTEM