Related ArticlesSemisynthesis of a segmental isotopically labeled protein splicing precursor: NMR evidence for an unusual peptide bond at the N-extein-intein junction.
Proc Natl Acad Sci U S A. 2004 Apr 27;101(17):6397-402
Authors: Romanelli A, Shekhtman A, Cowburn D, Muir TW
Protein splicing is a posttranslational autocatalytic process in which an intervening sequence, termed an intein, is removed from a host protein, the extein. Although we have a reasonable picture of the basic chemical steps in protein splicing, our knowledge of how these are catalyzed and regulated is less well developed. In the current study, a combination of NMR spectroscopy and segmental isotopic labeling has been used to study the structure of an active protein splicing precursor, corresponding to an N-extein fusion of the Mxe GyrA intein. The (1)J(NC') coupling constant for the (-1) scissile peptide bond at the N-extein-intein junction was found to be approximately 12 Hz, which indicates that this amide is highly polarized, perhaps because of nonplanarity. Additional mutagenesis and NMR studies indicate that conserved box B histidine residue is essential for catalysis of the first step of splicing and for maintaining the (-1) scissile bond in its unusual conformation. Overall, these studies support the "ground-state destabilization" model as part of the mechanism of catalysis.
Evidence from solid-state NMR for nonhelical conformations in the transmembrane domain of the amyloid precursor protein.
Evidence from solid-state NMR for nonhelical conformations in the transmembrane domain of the amyloid precursor protein.
Evidence from solid-state NMR for nonhelical conformations in the transmembrane domain of the amyloid precursor protein.
Biophys J. 2011 Feb 2;100(3):711-9
Authors: Lu JX, Yau WM, Tycko R
The amyloid precursor protein (APP) is subject to proteolytic processing by ?-secretase within neuronal membranes, leading to Alzheimer's disease-associated ?-amyloid peptide production by cleavage near the midpoint of the*single...
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Production of isotopically labeled heterologous proteins in non-E. coli prokaryotic and eukaryotic cells
Production of isotopically labeled heterologous proteins in non-E. coli prokaryotic and eukaryotic cells
Abstract The preparation of stable isotope-labeled proteins is necessary for the application of a wide variety of NMR methods, to study the structures and dynamics of proteins and protein complexes. The E. coli expression system is generally used for the production of isotope-labeled proteins, because of the advantages of ease of handling, rapid growth, high-level protein production, and low cost for isotope-labeling. However, many eukaryotic proteins are not functionally expressed...
[NMR paper] Simplification of protein NOESY spectra using bioorganic precursor synthesis and NMR
Simplification of protein NOESY spectra using bioorganic precursor synthesis and NMR spectral editing.
Related Articles Simplification of protein NOESY spectra using bioorganic precursor synthesis and NMR spectral editing.
J Am Chem Soc. 2004 May 5;126(17):5348-9
Authors: Lichtenecker R, Ludwiczek ML, Schmid W, Konrat R
A novel method is proposed for the analysis of protein NOEs in solution. In this approach, chemically synthesized precursor compounds for the amino acids valine, leucine, and isoleucine are used for amino acid specific labeling...
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[NMR paper] Overexpression and purification of isotopically labeled Escherichia coli MutH for NMR
Overexpression and purification of isotopically labeled Escherichia coli MutH for NMR studies.
Related Articles Overexpression and purification of isotopically labeled Escherichia coli MutH for NMR studies.
Protein Expr Purif. 2003 Jun;29(2):252-8
Authors: Dutta A, Rao BJ, Chary KV
MutH is one of the enzymes involved in the methyl directed -GATC-based DNA repair system. We report a significantly optimized protocol to prepare isotopically (15N and/or 13C) labeled MutH in minimal medium with high yields for NMR studies. Under the various...
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[NMR paper] Phosphorylation-induced structural changes in the amyloid precursor protein cytoplasm
Phosphorylation-induced structural changes in the amyloid precursor protein cytoplasmic tail detected by NMR.
Related Articles Phosphorylation-induced structural changes in the amyloid precursor protein cytoplasmic tail detected by NMR.
J Mol Biol. 2001 Mar 30;307(3):871-84
Authors: Ramelot TA, Nicholson LK
The cytoplasmic tail of the amyloid precursor protein (APPc) interacts with several cellular factors implicated in intracellular signaling or proteolytic production of amyloid beta peptide found in senile plaques of Alzheimer's disease...
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11-19-2010 08:32 PM
[NMR paper] Isotopically labeled bovine beta-lactoglobulin for NMR studies expressed in Pichia pa
Isotopically labeled bovine beta-lactoglobulin for NMR studies expressed in Pichia pastoris.
Related Articles Isotopically labeled bovine beta-lactoglobulin for NMR studies expressed in Pichia pastoris.
Protein Expr Purif. 1998 Oct;14(1):97-103
Authors: Denton H, Smith M, Husi H, Uhrin D, Barlow PN, Batt CA, Sawyer L
beta-Lactoglobulin (beta-Lg) is the major whey protein in ruminant milk and has been implicated in the irreversible denaturation of milk proteins and its associated poor processing behavior during heat treatment. In order to help...
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[NMR paper] The mitochondrial precursor protein apocytochrome c strongly influences the order of
The mitochondrial precursor protein apocytochrome c strongly influences the order of the headgroup and acyl chains of phosphatidylserine dispersions. A 2H and 31P NMR study.
Related Articles The mitochondrial precursor protein apocytochrome c strongly influences the order of the headgroup and acyl chains of phosphatidylserine dispersions. A 2H and 31P NMR study.
Biochemistry. 1990 Mar 6;29(9):2312-21
Authors: Jordi W, de Kroon AI, Killian JA, de Kruijff B
Deuterium and phosphorus nuclear magnetic resonance techniques were used to study the...
Semisynthesis of a segmental isotopically labeled protein splicing precursor: NMR evi
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