1gpx: Difference between revisions
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==Overview== | ==Overview== | ||
The Fe2S2 cluster of the ferredoxin putidaredoxin (Pdx) can be replaced by, a single gallium ion, giving rise to a colorless, diamagnetic protein in, which, apart from the metal binding site, the major structural features of, the native ferredoxin are conserved. The solution structure of the C85S, variant of gallium putidaredoxin (C85S GaPdx), in which a non-ligand, cysteine is replaced by a serine, has been determined via multidimensional, NMR methods using uniformly 15N, 13C labeled samples of C85S GaPdx., Stereospecific assignments of leucine and valine methyl resonances were, made using 13C, 1H HSQC spectra obtained with fractionally 13C-labeled, samples, and backbone dihedral angle restraints were obtained using a, combination of two-dimensional J-modulated 15N, 1H HSQC and, ... | The Fe2S2 cluster of the ferredoxin putidaredoxin (Pdx) can be replaced by, a single gallium ion, giving rise to a colorless, diamagnetic protein in, which, apart from the metal binding site, the major structural features of, the native ferredoxin are conserved. The solution structure of the C85S, variant of gallium putidaredoxin (C85S GaPdx), in which a non-ligand, cysteine is replaced by a serine, has been determined via multidimensional, NMR methods using uniformly 15N, 13C labeled samples of C85S GaPdx., Stereospecific assignments of leucine and valine methyl resonances were, made using 13C, 1H HSQC spectra obtained with fractionally 13C-labeled, samples, and backbone dihedral angle restraints were obtained using a, combination of two-dimensional J-modulated 15N, 1H HSQC and, three-dimensional (HN)CO(CO)NH experiments. A total of 1117 NOE-derived, distance restraints were used in the calculations, including 454 short, range (i-j < or = 3), 456 long range (i-j > or = 4) interresidue, restraints and 207 non-trivial intraresidue restraints. 97 phi and 55 chi, 1 angular restraints were also included in the calculation of a family of, 20 structures using a combined distance geometry-simulated annealing, protocol. Most regions of the protein are well defined in the, calculations, with an RMSD of 0.525 A for backbone atoms excluding the, metal binding loop (residues 34-48) and the last three C-terminal residues, (residues 103-106). Where comparison is possible, these regions show an, increase in dynamic behavior over the native protein, as does the loop, containing residues 74-76. Structural and dynamic differences between, native Pdx and GaPdx are discussed in relation to charge and packing of, the metal binding site. | ||
==About this Structure== | ==About this Structure== | ||
1GPX is a | 1GPX is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pseudomonas_putida Pseudomonas putida] with GA as [http://en.wikipedia.org/wiki/ligand ligand]. Structure known Active Site: GAL. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1GPX OCA]. | ||
==Reference== | ==Reference== | ||
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[[Category: gapdx c85s]] | [[Category: gapdx c85s]] | ||
''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on | ''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Mon Nov 5 14:13:31 2007'' | ||
Revision as of 15:08, 5 November 2007
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C85S GAPDX, NMR, 20 STRUCTURES
OverviewOverview
The Fe2S2 cluster of the ferredoxin putidaredoxin (Pdx) can be replaced by, a single gallium ion, giving rise to a colorless, diamagnetic protein in, which, apart from the metal binding site, the major structural features of, the native ferredoxin are conserved. The solution structure of the C85S, variant of gallium putidaredoxin (C85S GaPdx), in which a non-ligand, cysteine is replaced by a serine, has been determined via multidimensional, NMR methods using uniformly 15N, 13C labeled samples of C85S GaPdx., Stereospecific assignments of leucine and valine methyl resonances were, made using 13C, 1H HSQC spectra obtained with fractionally 13C-labeled, samples, and backbone dihedral angle restraints were obtained using a, combination of two-dimensional J-modulated 15N, 1H HSQC and, three-dimensional (HN)CO(CO)NH experiments. A total of 1117 NOE-derived, distance restraints were used in the calculations, including 454 short, range (i-j < or = 3), 456 long range (i-j > or = 4) interresidue, restraints and 207 non-trivial intraresidue restraints. 97 phi and 55 chi, 1 angular restraints were also included in the calculation of a family of, 20 structures using a combined distance geometry-simulated annealing, protocol. Most regions of the protein are well defined in the, calculations, with an RMSD of 0.525 A for backbone atoms excluding the, metal binding loop (residues 34-48) and the last three C-terminal residues, (residues 103-106). Where comparison is possible, these regions show an, increase in dynamic behavior over the native protein, as does the loop, containing residues 74-76. Structural and dynamic differences between, native Pdx and GaPdx are discussed in relation to charge and packing of, the metal binding site.
About this StructureAbout this Structure
1GPX is a Single protein structure of sequence from Pseudomonas putida with GA as ligand. Structure known Active Site: GAL. Full crystallographic information is available from OCA.
ReferenceReference
The solution structure of a gallium-substituted putidaredoxin mutant: GaPdx C85S., Pochapsky TC, Kuti M, Kazanis S, J Biomol NMR. 1998 Oct;12(3):407-15. PMID:9835048
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