Related ArticlesIsotope labeling strategies for analysis of an ion channel cytoplasmic domain by NMR spectroscopy.
Methods Mol Biol. 2013;998:289-300
Authors: Abarca-Heidemann K, Duchardt-Ferner E, Woehnert J, Rothberg BS
Abstract
As large, multimeric, integral membrane proteins, ion channels pose technical challenges to analysis by NMR spectroscopy. Here we present a strategy to overcome some of these technical hurdles, using a representative ion channel modulatory domain, the regulator of K(+) conductance (RCK) domain from a K(+) channel cloned from Thermoplasma volcanium. By introducing a mutation to limit the stoichiometry of the octameric RCK domain "gating ring" complex to its dimeric building block, NMR spectral resolution can be greatly improved. Here we present protocols for efficient production of highly deuterated, uniformly (15)N-labeled protein, as well as protein containing (15)N-labeling to specific amino acid types. These labeling strategies can be applied to improve spectral resolution and facilitate sequential resonance assignments.
Isotope labeling strategies for NMR studies of RNA
Isotope labeling strategies for NMR studies of RNA
Abstract The known biological functions of RNA have expanded in recent years and now include gene regulation, maintenance of sub-cellular structure, and catalysis, in addition to propagation of genetic information. As for proteins, RNA function is tightly correlated with structure. Unlike proteins, structural information for larger, biologically functional RNAs is relatively limited. NMR signal degeneracy, relaxation problems, and a paucity of long-range 1Hâ??1H dipolar contacts have limited the utility of traditional NMR approaches....
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01-09-2011 12:46 PM
Cell-free expression and stable isotope labelling strategies for membrane proteins
Cell-free expression and stable isotope labelling strategies for membrane proteins
Abstract Membrane proteins are highly underrepresented in the structural data-base and remain one of the most challenging targets for functional and structural elucidation. Their roles in transport and cellular communication, furthermore, often make over-expression toxic to their host, and their hydrophobicity and structural complexity make isolation and reconstitution a complicated task, especially in cases where proteins are targeted to inclusion bodies. The development of cell-free expression systems...
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01-09-2011 12:46 PM
[NMR paper] Site-selective labeling strategies for screening by NMR.
Site-selective labeling strategies for screening by NMR.
Related Articles Site-selective labeling strategies for screening by NMR.
Comb Chem High Throughput Screen. 2002 Dec;5(8):623-30
Authors: Weigelt J, Wikström M, Schultz J, van Dongen MJ
NMR based screening has become an important tool in the pharmaceutical industry. Methods that provide information on the location of small molecule binding sites on the surface of a drug target (e. g. SAR-by-NMR and related techniques) are of particular interest. In order to extend the applicability of...
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11-24-2010 08:58 PM
[NMR paper] New developments in isotope labeling strategies for protein solution NMR spectroscopy
New developments in isotope labeling strategies for protein solution NMR spectroscopy.
Related Articles New developments in isotope labeling strategies for protein solution NMR spectroscopy.
Curr Opin Struct Biol. 2000 Oct;10(5):585-92
Authors: Goto NK, Kay LE
The development of novel isotope labeling strategies for proteins has facilitated the study of the structure and dynamics of these molecules. In addition, the recent emergence of alternative methods of bacterial expression for obtaining isotopically labeled proteins permits the study of...
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11-19-2010 08:29 PM
[NMR paper] An advantage for use of isotope labeling and NMR chemical shifts to analyze the struc
An advantage for use of isotope labeling and NMR chemical shifts to analyze the structure of four homologous IgG-binding domains of staphylococcal protein A.
Related Articles An advantage for use of isotope labeling and NMR chemical shifts to analyze the structure of four homologous IgG-binding domains of staphylococcal protein A.
J Biochem Biophys Methods. 2000 Jan 3;42(1-2):35-47
Authors: Kikuchi J, Asakura T, Hasuda K, Ito T, Ohwaku K, Araki H, Williamson MP
Because of the complexity arising from the large molecular size and the amino acid...
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11-18-2010 09:15 PM
[NMR paper] Conformation of the cytoplasmic domain of phospholamban by NMR and CD.
Conformation of the cytoplasmic domain of phospholamban by NMR and CD.
Related Articles Conformation of the cytoplasmic domain of phospholamban by NMR and CD.
Mol Membr Biol. 1994 Oct-Dec;11(4):263-9
Authors: Hubbard JA, MacLachlan LK, Meenan E, Salter CJ, Reid DG, Lahouratate P, Humphries J, Stevens N, Bell D, Neville WA
Nuclear magnetic resonance (NMR) and circular dichroism (CD) spectroscopy have been used to characterize the conformation of the putative cytoplasmic domain of phospholamban (PLB), an oligomeric membrane-bound protein which...
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08-22-2010 03:29 AM
[KPWU blog] [IDP] Cytoplasmic Domain of the T Cell Receptor zeta
Cytoplasmic Domain of the T Cell Receptor zeta
Title:* The Intrinsically Disordered Cytoplasmic Domain of the T Cell Receptor ? Chain Binds to the Nef Protein of Simian Immunodeficiency Virus without a Disorder-to-Order Transition Authors: Alexander B. Sigalov, Walter M. Kim, Maria Saline and Lawrence J. Stern Journal: Biochemistry, 2008, 47 (49), pp 12942–12944 Not fully labeled in HSQChttp://stats.wordpress.com/b.gif?host=kpwu.wordpress.com&blog=76132&post=197&subd=kpwu&ref=&feed=1
Go to KPWU blog to read complete post.
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08-22-2010 01:33 AM
Cell-free expression and labeling strategies for a new decade in solid-state NMR.
Cell-free expression and labeling strategies for a new decade in solid-state NMR.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Cell-free expression and labeling strategies for a new decade in solid-state NMR.
N Biotechnol. 2010 Aug 3;
Authors: Abdine A, Verhoeven MA, Warschawski DE
Although solid-state NMR and cell-free expression have recently become standard methods in biology, the combination of the two is still at a very early stage of development. In this...
Isotope labeling strategies for analysis of an ion channel cytoplasmic domain by NMR spectroscopy.
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