Related ArticlesMonitoring the effects of antagonists on protein-protein interactions with NMR spectroscopy.
J Am Chem Soc. 2005 Sep 28;127(38):13220-6
Authors: D'Silva L, Ozdowy P, Krajewski M, Rothweiler U, Singh M, Holak TA
We describe an NMR method that directly monitors the influence of ligands on protein-protein interactions. For a two-protein interaction complex, the size of one component should be small enough (less than ca. 15 kDa) to provide a good quality (15)N((13)C) HSQC spectrum after (15)N((13)C) labeling. The size of the second unlabeled component should be large enough so that the molecular weight of the preformed complex is larger than ca. 40 kDa. When the smaller protein binds to a larger one, broadening of NMR resonances results in the disappearance of most of its cross-peaks in the HSQC spectrum. Addition of an antagonist that can dissociate the complex would restore the HSQC spectrum of the smaller component. The method directly shows whether an antagonist releases proteins in their wild-type folded states or whether it induces their denaturation, partial unfolding, or precipitation. We illustrate the method by studying lead compounds that have recently been reported to block the MDM2-p53 interaction. Activation of p53 in tumor cells by inhibiting its interaction with MDM2 offers new strategy for cancer therapy.
Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy.
Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy.
Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy.
Biochemistry. 2011 Sep 26;
Authors: Wang Q, Zhuravleva A, Gierasch LM
Abstract
Biology relies on functional interplay of proteins in the crowded and heterogeneous environment inside cells, and functional protein interactions are often weak and transient. Thus, methods are needed that preserve these interactions...
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Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy.
Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy.
Exploring weak, transient protein-protein interactions in crowded in vivo environments by in-cell NMR spectroscopy.
Biochemistry. 2011 Sep 26;
Authors: Wang Q, Zhuravleva A, Gierasch LM
Abstract
Biology relies on functional interplay of proteins in the crowded and heterogeneous environment inside cells, and functional protein interactions are often weak and transient. Thus, methods are needed that preserve these...
[NMR paper] New structural insights into carbohydrate-protein interactions from NMR spectroscopy.
New structural insights into carbohydrate-protein interactions from NMR spectroscopy.
Related Articles New structural insights into carbohydrate-protein interactions from NMR spectroscopy.
Curr Opin Struct Biol. 2003 Oct;13(5):646-53
Authors: Kogelberg H, Solís D, Jiménez-Barbero J
Recently developed NMR methods have been applied to discover carbohydrate ligands for proteins and to identify their binding epitopes. The structural details of carbohydrate-protein complexes have also been examined by NMR, providing site-specific information on the...
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11-24-2010 09:16 PM
[NMR paper] Transient protein interactions studied by NMR spectroscopy: the case of cytochrome C
Transient protein interactions studied by NMR spectroscopy: the case of cytochrome C and adrenodoxin.
Related Articles Transient protein interactions studied by NMR spectroscopy: the case of cytochrome C and adrenodoxin.
Biochemistry. 2003 Jun 17;42(23):7068-76
Authors: Worrall JA, Reinle W, Bernhardt R, Ubbink M
The interaction between yeast iso-1-cytochrome c (C102T) and two forms of bovine adrenodoxin, the wild type and a truncated form comprising residues 4-108, has been investigated using a combination of one- and two-dimensional...
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[NMR paper] Facile detection of protein-protein interactions by one-dimensional NMR spectroscopy.
Facile detection of protein-protein interactions by one-dimensional NMR spectroscopy.
Related Articles Facile detection of protein-protein interactions by one-dimensional NMR spectroscopy.
Biochemistry. 2003 Mar 18;42(10):2774-80
Authors: Araç D, Murphy T, Rizo J
Two methods for detecting protein-protein interactions in solution using one-dimensional (1D) NMR spectroscopy are described. Both methods rely on measurement of the intensity of the strongest methyl resonance (SMR), which for most proteins is observed at 0.8-0.9 ppm. The severe...
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[NMR paper] NMR monitoring of accumulation and folding of 15N-labeled protein overexpressed in Pi
NMR monitoring of accumulation and folding of 15N-labeled protein overexpressed in Pichia pastoris.
Related Articles NMR monitoring of accumulation and folding of 15N-labeled protein overexpressed in Pichia pastoris.
Protein Expr Purif. 2001 Jul;22(2):318-24
Authors: de Lamotte F, Boze H, Blanchard C, Klein C, Moulin G, Gautier MF, Delsuc MA
Postgenomic studies have led to an increasing demand for isotope-labeled proteins. We present a method for producing large quantities of truly native (15)N-labeled protein. Based on the secretion...
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11-19-2010 08:44 PM
[NMR paper] Probing carbohydrate-protein interactions by high-resolution NMR spectroscopy.
Probing carbohydrate-protein interactions by high-resolution NMR spectroscopy.
Related Articles Probing carbohydrate-protein interactions by high-resolution NMR spectroscopy.
Adv Exp Med Biol. 1998;435:29-38
Authors: Homans SW, Field RA, Milton MJ, Probert M, Richardson JM