[NMR paper] A Facile method for expression and purification of (15)N isotope-labeled human Alzheimer's ?-amyloid peptides from E. coli for NMR-based structural analysis.
A Facile method for expression and purification of (15)N isotope-labeled human Alzheimer's ?-amyloid peptides from E. coli for NMR-based structural analysis.
Related ArticlesA Facile method for expression and purification of (15)N isotope-labeled human Alzheimer's ?-amyloid peptides from E. coli for NMR-based structural analysis.
Protein Expr Purif. 2015 Jul 28;
Authors: Sharma SC, Armand T, Aurelia Ball K, Chen A, Pelton JG, Wemmer DE, Head-Gordon T
Abstract
Alzheimer's disease (AD) is a progressive neurodegenerative disease affecting millions of people worldwide. AD is characterized by the presence of extracellular plaques composed of aggregated/oligomerized ?-amyloid peptides with A?42 peptide representing a major isoform in the senile plaques. Given the pathological significance of A?42 in the progression of AD, there is considerable interest in understanding the structural ensembles for soluble monomer and oligomeric forms of A?42. This report describes an efficient method to express and purify high quality (15)N isotope-labeled A?42 for structural studies by NMR. The protocol involves utilization of an auto induction system with (15)N isotope labeled medium, for high-level expression of A?42 as a fusion with IFABP. After the over-expression of the (15)N isotope-labeled IFABP-A?42 fusion protein in the inclusion bodies, pure (15)N isotope-labeled A?42 peptide is obtained following a purification method that is streamlined and improved from the method originally developed for the isolation of unlabeled A?42 peptide (Protein Expr. Purif. 66 (2009) 107-112). We obtain a final yield of ~6 mg/L culture for (15)N isotope-labeled A?42 peptide. Mass spectrometry and (1)H-(15)N HSQC spectra of monomeric A?42 peptide validate the uniform incorporation of the isotopic label. The method described here is equally applicable for the uniform isotope labeling with (15)N and (13)C in A?42 peptide as well as its other variants including any A?42 peptide mutants.
PMID: 26231074 [PubMed - as supplied by publisher]
[NMR paper] Preparation of uniformly isotope labeled KcsA for solid state NMR: Expression, purification, reconstitution into liposomes and functional assay.
Preparation of uniformly isotope labeled KcsA for solid state NMR: Expression, purification, reconstitution into liposomes and functional assay.
Preparation of uniformly isotope labeled KcsA for solid state NMR: Expression, purification, reconstitution into liposomes and functional assay.
Protein Expr Purif. 2013 Aug 1;
Authors: Bhate MP, Wylie BJ, Thompson A, Tian L, Nimigean C, McDermott AE
Abstract
We report the expression, purification, liposome reconstitution and functional validation of uniformly (13)C and (15)N isotope labeled...
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[NMR paper] Structural analysis of the pyroglutamate-modified isoform of the Alzheimer's disease-related amyloid-? using NMR spectroscopy.
Structural analysis of the pyroglutamate-modified isoform of the Alzheimer's disease-related amyloid-? using NMR spectroscopy.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--media.wiley.com-assets-2250-98-WileyOnlineLibrary-Button_120x27px_FullText.gif Related Articles Structural analysis of the pyroglutamate-modified isoform of the Alzheimer's disease-related amyloid-? using NMR spectroscopy.
J Pept Sci. 2012 Nov;18(11):691-5
Authors: Sun N, Hartmann R, Lecher J, Stoldt M, Funke SA, Gremer L, Ludwig HH, Demuth HU, Kleinschmidt M,...
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Expression, purification and preliminary NMR characterization of isotopically labeled wild-type human heterotrimeric G protein ?i1
Expression, purification and preliminary NMR characterization of isotopically labeled wild-type human heterotrimeric G protein ?i1
August 2012
Publication year: 2012
Source:Protein Expression and Purification, Volume 84, Issue 2</br>
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Molecular-level investigation of proteins is increasingly important to researchers trying to understand the mechanisms of signal transmission. Heterotrimeric G proteins control the activation of many critical signal transmission cascades and are also implicated in numerous diseases. As part of a longer-term investigation of...
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Expression, purification, and reconstitution of the transmembrane domain of the human amyloid precursor protein for NMR studies.
Expression, purification, and reconstitution of the transmembrane domain of the human amyloid precursor protein for NMR studies.
Expression, purification, and reconstitution of the transmembrane domain of the human amyloid precursor protein for NMR studies.
Protein Expr Purif. 2011 Aug 31;
Authors: Chen W, Gamache E, Richardson D, Du Z, Wang C
Abstract
Alzheimer's disease (AD) is the most common type of dementia in elderly people. Senile plaques, a pathologic hallmark of AD, are composed of amyloid ? peptide (A?). A? aggregation produces...
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09-13-2011 08:27 PM
Expression and purification of (15)N- and (13)C-isotope labeled 40-residue human Alzheimer's ?-amyloid peptide for NMR-based structural analysis.
Expression and purification of (15)N- and (13)C-isotope labeled 40-residue human Alzheimer's ?-amyloid peptide for NMR-based structural analysis.
Expression and purification of (15)N- and (13)C-isotope labeled 40-residue human Alzheimer's ?-amyloid peptide for NMR-based structural analysis.
Protein Expr Purif. 2011 May 27;
Authors: Long F, Cho W, Ishii Y
Amyloid fibrils of Alzheimer's ?-amyloid peptide (A?) are a primary component of amyloid plaques, a hallmark of Alzheimer's disease (AD). Enormous attention has been given to the structural...
[NMR paper] Expression and purification of a recombinant peptide from the Alzheimer's beta-amyloi
Expression and purification of a recombinant peptide from the Alzheimer's beta-amyloid protein for solid-state NMR.
Related Articles Expression and purification of a recombinant peptide from the Alzheimer's beta-amyloid protein for solid-state NMR.
Protein Expr Purif. 2005 Jul;42(1):200-10
Authors: Sharpe S, Yau WM, Tycko R
Fibrillar protein aggregates contribute to the pathology of a number of disease states. To facilitate structural studies of these amyloid fibrils by solid-state NMR, efficient methods for the production of milligram...
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11-24-2010 11:14 PM
An economical method for producing stable-isotope labeled proteins by the E. coli cel
An economical method for producing stable-isotope labeled proteins by the E. coli cell-free system
Abstract Improvement of the cell-free protein synthesis system (CF) over the past decade have made it one of the most powerful protein production methods. The CF approach is especially useful for stable-isotope (SI) labeling of proteins for NMR analysis. However, it is less popular than expected, partly because the SI-labeled amino acids used for SI labeling by the CF are too expensive. In the present study, we developed a simple and inexpensive method for producing an SI-labeled protein...
A Facile method for expression and purification of (15)N isotope-labeled human Alzheimer's ?-amyloid peptides from E. coli for NMR-based structural analysis.
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