1gl8
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SOLUTION STRUCTURE OF THIOREDOXIN M FROM SPINACH, OXIDIZED FORM
OverviewOverview
Proton NMR spectral resonances of thioredoxin m from spinach have been, assigned, and its solution structure has been determined on the basis of, 1156 nuclear Overhauser effect- (NOE-) derived distance constraints by, using restrained molecular dynamics calculations. The average pairwise, root-mean-square deviation (RMSD) for the 25 best NMR structures for the, backbone was 1.0 +/- 0.1, when the structurally well-defined residues were, considered. The N- and C-terminal segments (1-13 and 118-119) and residues, 41-49, comprising the active site, are highly disordered. At the time of, concluding this work, a crystal structure of this protein was reported, in, which thioredoxin m was found to crystallize as noncovalent dimers., Although the solution and crystal structures are very similar, no evidence, was found about the existence of dimers in solution, thus confirming that, dimerization is not needed for the regulatory activity of thioredoxin m., The spinach thioredoxin m does not unfold by heat in the range 25-85, degrees C, as revealed by thermal circular dichroic (CD) measurements., However, its unfolding free energy (9.1 +/- 0.8 kcal mol(-1), at pH 5.3, and 25 degrees C) could be determined by extrapolating the free energy, values obtained at different concentrations of guanidinium chloride, (GdmCl). The folding-unfolding process is two-state as indicated by the, coincidence of the CD denaturation curves obtained at far and near UV. The, H/D exchange behavior of backbone amide protons was analyzed. The, slowest-exchanging protons, requiring a global-unfolding mechanism in, order to exchange, are those from beta2, beta3, and beta4, the central, strands of the beta-sheet, which constitute the main element of the core, of the protein. The free energies obtained from exchange measurements of, protons belonging to the alpha-helices are lower than those derived from, GdmCl denaturation studies, indicating that those protons exchange by, local-unfolding mechanisms.
About this StructureAbout this Structure
1GL8 is a Single protein structure of sequence from Spinacia oleracea. Full crystallographic information is available from OCA.
ReferenceReference
Three-dimensional solution structure and stability of thioredoxin m from spinach., Neira JL, Gonzalez C, Toiron C, de Prat-Gay G, Rico M, Biochemistry. 2001 Dec 18;40(50):15246-56. PMID:11735407
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