Expression and purification of a recombinant amyloidogenic peptide from transthyretin for solid-state NMR spectroscopy

Philippe S. Nadaud; Mohosin Sarkar; Bo Wu; Cait E. MacPhee; Thomas J. Magliery; Christopher P. Jaroniec

doi:10.1016/j.pep.2009.09.017

Expression and purification of a recombinant amyloidogenic peptide from transthyretin for solid-state NMR spectroscopy

Philippe S. Nadaud, Mohosin Sarkar, Bo Wu, Cait E. MacPhee, Thomas J. Magliery, Christopher P. Jaroniec

School of Physics and Astronomy

Research output: Contribution to journal › Article › peer-review

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
Pages (from-to)	101-108
Number of pages	8
Journal	Protein Expression and Purification
Volume	70
Issue number	1
DOIs	https://doi.org/10.1016/j.pep.2009.09.017
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

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10.1016/j.pep.2009.09.017

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title = "Expression and purification of a recombinant amyloidogenic peptide from transthyretin for solid-state NMR spectroscopy",

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.",

keywords = "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",

author = "Nadaud, {Philippe S.} and Mohosin Sarkar and Bo Wu and MacPhee, {Cait E.} and Magliery, {Thomas J.} and Jaroniec, {Christopher P.}",

year = "2010",

month = mar,

doi = "10.1016/j.pep.2009.09.017",

language = "English",

volume = "70",

pages = "101--108",

journal = "Protein Expression and Purification",

issn = "1046-5928",

publisher = "Academic Press",

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T1 - Expression and purification of a recombinant amyloidogenic peptide from transthyretin for solid-state NMR spectroscopy

AU - Nadaud, Philippe S.

AU - Sarkar, Mohosin

AU - Wu, Bo

AU - MacPhee, Cait E.

AU - Magliery, Thomas J.

AU - Jaroniec, Christopher P.

PY - 2010/3

Y1 - 2010/3

N2 - 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.

AB - 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.

KW - Fusion protein

KW - GB1

KW - Recombinant human transthyretin 105-115

KW - Escherichia coli BL21(DE3)

KW - Ni2+ affinity chromatography

KW - Amyloid fibrils

KW - Solid-state NMR spectroscopy

KW - ALPHA-SYNUCLEIN FIBRILS

KW - BETA-SHEET STRUCTURE

KW - HUMAN PRION PROTEIN

KW - ANGLE-SPINNING NMR

KW - ESCHERICHIA-COLI

KW - HIGH-RESOLUTION

KW - MOLECULAR-CONFORMATION

KW - STRUCTURAL MODEL

KW - RESIDUES

KW - SYSTEM

UR - http://www.scopus.com/inward/record.url?scp=75349113179&partnerID=8YFLogxK

U2 - 10.1016/j.pep.2009.09.017

DO - 10.1016/j.pep.2009.09.017

M3 - Article

SN - 1046-5928

VL - 70

SP - 101

EP - 108

JO - Protein Expression and Purification

JF - Protein Expression and Purification

IS - 1

ER -

Expression and purification of a recombinant amyloidogenic peptide from transthyretin for solid-state NMR spectroscopy

Abstract

Keywords / Materials (for Non-textual outputs)

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