2rlz
From Proteopedia
(New page: '''Unreleased structure''' The entry 2rlz is ON HOLD until Paper Publication Authors: Loquet, A., Bardiaux, B., Gardiennet, C., Blanchet, C., Baldus, M., Nilges, M., Malliavin, T., Bock...) |
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| - | + | [[Image:2rlz.jpg|left|200px]] | |
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| + | You may change the PDB parameter (which sets the PDB file loaded into the applet) | ||
| + | or the SCENE parameter (which sets the initial scene displayed when the page is loaded), | ||
| + | or leave the SCENE parameter empty for the default display. | ||
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| + | {{STRUCTURE_2rlz| PDB=2rlz | SCENE= }} | ||
| - | + | '''Solid-State MAS NMR structure of the dimer Crh''' | |
| - | Description: Solid-State MAS NMR structure of the dimer Crh | ||
| + | ==Overview== | ||
| + | In a wide variety of proteins, insolubility presents a challenge to structural biology, as X-ray crystallography and liquid-state NMR are unsuitable. Indeed, no general approach is available as of today for studying the three-dimensional structures of membrane proteins and protein fibrils. We here demonstrate, at the example of the microcrystalline model protein Crh, how high-resolution 3D structures can be derived from magic-angle spinning solid-state NMR distance restraints for fully labeled protein samples. First, we show that proton-mediated rare-spin correlation spectra, as well as carbon-13 spin diffusion experiments, provide enough short, medium, and long-range structural restraints to obtain high-resolution structures of this 2 x 10.4 kDa dimeric protein. Nevertheless, the large number of 13C/15N spins present in this protein, combined with solid-state NMR line widths of about 0.5-1 ppm, induces substantial ambiguities in resonance assignments, preventing 3D structure determination by using distance restraints uniquely assigned on the basis of their chemical shifts. In the second part, we thus demonstrate that an automated iterative assignment algorithm implemented in a dedicated solid-state NMR version of the program ARIA permits to resolve the majority of ambiguities and to calculate a de novo 3D structure from highly ambiguous solid-state NMR data, using a unique fully labeled protein sample. We present, using distance restraints obtained through the iterative assignment process, as well as dihedral angle restraints predicted from chemical shifts, the 3D structure of the fully labeled Crh dimer refined at a root-mean-square deviation of 1.33 A. | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Jun | + | ==About this Structure== |
| + | 2RLZ is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2RLZ OCA]. | ||
| + | |||
| + | ==Reference== | ||
| + | 3D structure determination of the Crh protein from highly ambiguous solid-state NMR restraints., Loquet A, Bardiaux B, Gardiennet C, Blanchet C, Baldus M, Nilges M, Malliavin T, Bockmann A, J Am Chem Soc. 2008 Mar 19;130(11):3579-89. Epub 2008 Feb 20. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/18284240 18284240] | ||
| + | [[Category: Bacillus subtilis]] | ||
| + | [[Category: Single protein]] | ||
| + | [[Category: Baldus, M.]] | ||
| + | [[Category: Bardiaux, B.]] | ||
| + | [[Category: Blanchet, C.]] | ||
| + | [[Category: Bockmann, A.]] | ||
| + | [[Category: Gardiennet, C.]] | ||
| + | [[Category: Loquet, A.]] | ||
| + | [[Category: Malliavin, T.]] | ||
| + | [[Category: Nilges, M.]] | ||
| + | [[Category: Dimer]] | ||
| + | [[Category: Domain-swap]] | ||
| + | [[Category: Ma]] | ||
| + | [[Category: Phosphorylation]] | ||
| + | [[Category: Solid-state nmr]] | ||
| + | [[Category: Transport protein]] | ||
| + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Jun 18 12:04:19 2008'' | ||
Revision as of 09:04, 18 June 2008
Solid-State MAS NMR structure of the dimer Crh
Overview
In a wide variety of proteins, insolubility presents a challenge to structural biology, as X-ray crystallography and liquid-state NMR are unsuitable. Indeed, no general approach is available as of today for studying the three-dimensional structures of membrane proteins and protein fibrils. We here demonstrate, at the example of the microcrystalline model protein Crh, how high-resolution 3D structures can be derived from magic-angle spinning solid-state NMR distance restraints for fully labeled protein samples. First, we show that proton-mediated rare-spin correlation spectra, as well as carbon-13 spin diffusion experiments, provide enough short, medium, and long-range structural restraints to obtain high-resolution structures of this 2 x 10.4 kDa dimeric protein. Nevertheless, the large number of 13C/15N spins present in this protein, combined with solid-state NMR line widths of about 0.5-1 ppm, induces substantial ambiguities in resonance assignments, preventing 3D structure determination by using distance restraints uniquely assigned on the basis of their chemical shifts. In the second part, we thus demonstrate that an automated iterative assignment algorithm implemented in a dedicated solid-state NMR version of the program ARIA permits to resolve the majority of ambiguities and to calculate a de novo 3D structure from highly ambiguous solid-state NMR data, using a unique fully labeled protein sample. We present, using distance restraints obtained through the iterative assignment process, as well as dihedral angle restraints predicted from chemical shifts, the 3D structure of the fully labeled Crh dimer refined at a root-mean-square deviation of 1.33 A.
About this Structure
2RLZ is a Single protein structure of sequence from Bacillus subtilis. Full crystallographic information is available from OCA.
Reference
3D structure determination of the Crh protein from highly ambiguous solid-state NMR restraints., Loquet A, Bardiaux B, Gardiennet C, Blanchet C, Baldus M, Nilges M, Malliavin T, Bockmann A, J Am Chem Soc. 2008 Mar 19;130(11):3579-89. Epub 2008 Feb 20. PMID:18284240 Page seeded by OCA on Wed Jun 18 12:04:19 2008
