Related ArticlesDifferential isotype labeling strategy for determining the structure of myristoylated recoverin by NMR spectroscopy.
J Biomol NMR. 1998 Feb;11(2):135-52
Authors: Tanaka T, Ames JB, Kainosho M, Stryer L, Ikura M
The three-dimensional solution structure of recombinant bovine myristoylated recoverin in the Ca(2+)-free state has been refined using an array of isotope-assisted multidimensional heteronuclear NMR techniques. In some experiments, the myristoyl group covalently attached to the protein N-terminus was labeled with C and the protein was unlabeled or vice versa; in others, both were C-labeled. This differential labeling strategy was essential for structural refinement and can be applied to other acylated proteins. Stereospecific assignments of 41 pairs of beta-methylene protons and 48 methyl groups of valine and leucine were included in the structure refinement. The refined structure was constructed using a total of 3679 experimental NMR restraints, comprising 3242 approximate interproton distance restraints (including 153 between the myristoyl group and the polypeptide), 140 distance restraints for 70 backbone hydrogen bonds, and 297 torsion angle restraints. The atomic rms deviations about the average minimized coordinate positions for the secondary structure region of the N-terminal and C-terminal domains are 0.44 +/- 0.07 and 0.55 +/- 0.18 A for backbone atoms, and the 1.09 +/- 0.07 and 1.10 +/- 0.15 A for all heavy atoms, respectively. The refined structure allows for a detailed analysis of the myristoyl binding pocket. The myristoyl group is in a slightly bent conformation: the average distance between C1 and C14 atoms of the myristoyl group is 14.6 A. Hydrophobic residues Leu28, Trp31, and Tyr32 from a cluster that interacts with the front end of the myristoyl (C1-C8), whereas residues Phe49, Phe56, Tyr86, Val87, and Leu90 interact with the tail end (C9-C14). The relatively deep hydrophobic pocket that binds the myristoyl group (C14:0) could also accommodate other naturally occurring acyl groups such as C12:0, C14:1, C14:2 chains.
A segmental labeling strategy for unambiguous determination of domainā??domain interactions of large multi-domain proteins
A segmental labeling strategy for unambiguous determination of domainā??domain interactions of large multi-domain proteins
Abstract NMR structural determination of large multi-domain proteins is a challenging task due to significant spectral overlap with a particular difficulty in unambiguous identification of domainā??domain interactions. Segmental labeling is a NMR strategy that allows for isotopically labeling one domain and leaves the other domain unlabeled. This significantly simplifies spectral overlaps and allows for quick identification of domainā??domain interaction. Here, a...
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A novel strategy for NMR resonance assignment and protein structure determination
A novel strategy for NMR resonance assignment and protein structure determination
Abstract The quality of protein structures determined by nuclear magnetic resonance (NMR) spectroscopy is contingent on the number and quality of experimentally-derived resonance assignments, distance and angular restraints. Two key features of protein NMR data have posed challenges for the routine and automated structure determination of small to medium sized proteins; (1) spectral resolution ā?? especially of crowded nuclear Overhauser effect spectroscopy (NOESY) spectra, and (2) the reliance on a...
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12-18-2010 01:31 AM
A novel strategy for NMR resonance assignment and protein structure determination.
A novel strategy for NMR resonance assignment and protein structure determination.
A novel strategy for NMR resonance assignment and protein structure determination.
J Biomol NMR. 2010 Dec 14;
Authors: Lemak A, Gutmanas A, Chitayat S, Karra M, Farčs C, Sunnerhagen M, Arrowsmith CH
The quality of protein structures determined by nuclear magnetic resonance (NMR) spectroscopy is contingent on the number and quality of experimentally-derived resonance assignments, distance and angular restraints. Two key features of protein NMR data have posed...
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12-17-2010 11:23 AM
A simple strategy for 13C,1H labeling at the Ile-Ī³2 methyl position in highly deuter
A simple strategy for 13C,1H labeling at the Ile-Ī³2 methyl position in highly deuterated proteins
Abstract A straightforward approach for the production of highly deuterated proteins labeled with 13C and 1H at Ile-Ī³2 methyl positions is described. The utility of the methodology is illustrated with an application involving the half proteasome (360 kDa). High quality 2D Ile 13CĪ³2,1HĪ³2 HMQC data sets, exploiting the methyl-TROSY principle, are recorded with excellent sensitivity and resolution, that compare favorably with Ile 13CĪ“1,1HĪ“1 spectra. This labeling scheme adds to a growing...
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Postdoc position in solid-state NMR: Determining biomolecular structure at the interf
Postdoc position in solid-state NMR: Determining biomolecular structure at the interface of structural and cellular biology
Description of the Faculty / Research group
The Faculty of Science consists of six departments: Biology, Pharmaceutical Sciences, Information and Computing Sciences, Physics and Astronomy, Mathematics and Chemistry. The Faculty is home to 3500 students and nearly 2000 staff and is internationally renowned for the quality of its research.
The NMR Research Group is part of the Chemistry Department (Universiteit Utrecht fac. Scheikunde) and...
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[NMR paper] Secondary structure of myristoylated recoverin determined by three-dimensional hetero
Secondary structure of myristoylated recoverin determined by three-dimensional heteronuclear NMR: implications for the calcium-myristoyl switch.
Related Articles Secondary structure of myristoylated recoverin determined by three-dimensional heteronuclear NMR: implications for the calcium-myristoyl switch.
Biochemistry. 1994 Sep 6;33(35):10743-53
Authors: Ames JB, Tanaka T, Stryer L, Ikura M
Recoverin, a new member of the EF-hand superfamily, serves as a Ca2+ sensor in vision. A myristoyl or related N-acyl group is covalently attached at its...
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08-22-2010 03:29 AM
[NMR paper] NMR strategy for determining Xaa-Pro peptide bond configurations in proteins: mutants
NMR strategy for determining Xaa-Pro peptide bond configurations in proteins: mutants of staphylococcal nuclease with altered configuration at proline-117.
Related Articles NMR strategy for determining Xaa-Pro peptide bond configurations in proteins: mutants of staphylococcal nuclease with altered configuration at proline-117.
Biochemistry. 1993 Nov 9;32(44):11810-8
Authors: Hinck AP, Eberhardt ES, Markley JL
A general approach has been developed for configurational analysis (cis or trans) of Xaa-Pro peptide bonds in proteins. This approach,...
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[NMR paper] A calculation strategy for the structure determination of symmetric dimers by 1H NMR.
A calculation strategy for the structure determination of symmetric dimers by 1H NMR.
Related Articles A calculation strategy for the structure determination of symmetric dimers by 1H NMR.
Proteins. 1993 Nov;17(3):297-309
Authors: Nilges M
The structure determination of symmetric dimers by NMR is impeded by the ambiguity of inter- and intramonomer NOE crosspeaks. In this paper, a calculation strategy is presented that allows the calculation of dimer structures without resolving the ambiguity by additional experiments (like asymmetric...