Publications
62 results found
Antcliffe D, Jimenez B, Veselkov K, et al., 2014, DIAGNOSING PNEUMONIA ON THE INTENSIVE CARE UNIT WITH SERUM <SUP>1</SUP>H NMR SPECTROSCOPY, 27th Annual Congress of the European-Society-of-Intensive-Care-Medicine (ESICM), Publisher: SPRINGER, Pages: S237-S238, ISSN: 0342-4642
Bray R, Jimenez B, MacIntyre D, et al., 2014, 1H-NMR BASED URINE METABONOMICS AS A NOVEL APPROACH TO OVERACTIVE BLADDER SYNDROME, 44th Annual Meeting of the International-Continence-Society (ICS), Publisher: WILEY-BLACKWELL, Pages: 923-924, ISSN: 0733-2467
Bray R, Jimenez B, MacIntyre D, et al., 2014, IS METABOLOMIC PROFILING USEFUL IN THE ASSESMENT OF WOMEN WITH LOWER URINARY TRACT SYMPTOMS?, 44th Annual Meeting of the International-Continence-Society (ICS), Publisher: WILEY-BLACKWELL, Pages: 926-927, ISSN: 0733-2467
Bray R, Jimenez B, MacIntyre D, et al., 2014, HIPPURIC ACID: A BIOMARKER FOR BLADDER PAIN SYNDROME, 44th Annual Meeting of the International-Continence-Society (ICS), Publisher: WILEY-BLACKWELL, Pages: 796-797, ISSN: 0733-2467
- Author Web Link
- Cite
- Citations: 1
Bray R, Jimenez B, MacIntyre DA, et al., 2014, 1H-NUCLEAR MAGNETIC RESONANCE URINARY METABOLOMICS IN OVERACTIVE BLADDER: DO NOCTURIA AND URGENCY HAVE SIMILAR METABOLIC PROFILES?, Publisher: SPRINGER LONDON LTD, Pages: S94-S95, ISSN: 0937-3462
Mirnezami R, Jimenez B, Li JV, et al., 2014, Rapid Diagnosis and Staging of Colorectal Cancer via High-Resolution Magic Angle Spinning Nuclear Magnetic Resonance (HR-MAS NMR) Spectroscopy of Intact Tissue Biopsies, Annals of Surgery, Vol: 259, Pages: 1138-1149, ISSN: 0003-4932
Objective: To develop novel metabolite-based models for diagnosis and staging in colorectal cancer (CRC) using high-resolution magic angle spinning nuclear magnetic resonance (HR-MAS NMR) spectroscopy.Background: Previous studies have demonstrated that cancer cells harbor unique metabolic characteristics relative to healthy counterparts. This study sought to characterize metabolic properties in CRC using HR-MAS NMR spectroscopy.Methods: Between November 2010 and January 2012, 44 consecutive patients with confirmed CRC were recruited to a prospective observational study. Fresh tissue samples were obtained from center of tumor and 5 cm from tumor margin from surgical resection specimens. Samples were run in duplicate where tissue volume permitted to compensate for anticipated sample heterogeneity. Samples were subjected to HR-MAS NMR spectroscopic profiling and acquired spectral data were imported into SIMCA and MATLAB statistical software packages for unsupervised and supervised multivariate analysis.Results: A total of 171 spectra were acquired (center of tumor, n = 88; 5 cm from tumor margin, n = 83). Cancer tissue contained significantly increased levels of lactate (P < 0.005), taurine (P < 0.005), and isoglutamine (P < 0.005) and decreased levels of lipids/triglycerides (P < 0.005) relative to healthy mucosa (R2Y = 0.94; Q2Y = 0.72; area under the curve, 0.98). Colon cancer samples (n = 49) contained higher levels of acetate (P < 0.005) and arginine (P < 0.005) and lower levels of lactate (P < 0.005) relative to rectal cancer samples (n = 39). In addition unique metabolic profiles were observed for tumors of differing T-stage.Conclusions: HR-MAS NMR profiling demonstrates cancer-specific metabolic signatures in CRC and reveals metabolic differences between colonic and rectal cancers. In addition, this approach reveals that tumor metabolism undergoes modification during local tumor advancement, offering potential in future staging and therapeu
MacIntyre DA, Lee YS, Marchesi J, et al., 2014, Preterm Premature Rupture of Membranes (PPROM) Is Associated with Disparate Vaginal Microbiomes That Are Correlated with Specific Urinary Metabolic Profiles, 61st Annual Scientific Meeting of the Society-for-Gynecologic-Investigation (SGI), Publisher: SAGE PUBLICATIONS INC, Pages: 246A-246A, ISSN: 1933-7191
Jimenez B, Drymousis P, Kinross J, et al., 2014, Metabonomic Profiling: A Novel Approach in Neuroendocrine Neoplasias, 11th Annual ENETS Conference for the Diagnosis and Treatment of Neuroendocrine Tumor Disease, Publisher: KARGER, Pages: 227-227, ISSN: 0028-3835
Kinross JM, Drymousis P, Jimenez B, et al., 2013, Metabonomic profiling: A novel approach in neuroendocrine neoplasias, SURGERY, Vol: 154, Pages: 1185-1192, ISSN: 0039-6060
- Author Web Link
- Cite
- Citations: 24
Jimenez B, Mirnezami R, Kinross J, et al., 2013, <SUP>1</SUP>H HR-MAS NMR Spectroscopy of Tumor-Induced Local Metabolic "Field-Effects" Enables Colorectal Cancer Staging and Prognostication, JOURNAL OF PROTEOME RESEARCH, Vol: 12, Pages: 959-968, ISSN: 1535-3893
- Author Web Link
- Cite
- Citations: 88
Wong A, Jimenez B, Li X, et al., 2012, Evaluation of High Resolution Magic-Angle Coil Spinning NMR Spectroscopy for Metabolic Profiling of Nanoliter Tissue Biopsies, ANALYTICAL CHEMISTRY, Vol: 84, Pages: 3843-3848, ISSN: 0003-2700
- Author Web Link
- Open Access Link
- Cite
- Citations: 34
Jiménez B, Ugwu F, Zhao R, et al., 2012, Structure of minimal tetratricopeptide repeat domain protein Tah1 reveals mechanism of its interaction with Pih1 and Hsp90., J Biol Chem, Vol: 287, Pages: 5698-5709
Tah1 and Pih1 are novel Hsp90 interactors. Tah1 acts as a cofactor of Hsp90 to stabilize Pih1. In yeast, Hsp90, Tah1, and Pih1 were found to form a complex that is required for ribosomal RNA processing through their effect on box C/D small nucleolar ribonucleoprotein assembly. Tah1 is a minimal tetratricopeptide repeat protein of 111 amino acid residues that binds to the C terminus of the Hsp90 molecular chaperone, whereas Pih1 consists of 344 residues of unknown fold. The NMR structure of Tah1 has been solved, and this structure shows the presence of two tetratricopeptide repeat motifs followed by a C helix and an unstructured region. The binding of Tah1 to Hsp90 is mediated by the EEVD C-terminal residues of Hsp90, which bind to a positively charged channel formed by Tah1. Five highly conserved residues, which form a two-carboxylate clamp that tightly interacts with the ultimate Asp-0 residue of the bound peptide, are also present in Tah1. Tah1 was found to bind to the C terminus of Pih1 through the C helix and the unstructured region. The C terminus of Pih1 destabilizes the protein in vitro and in vivo, whereas the binding of Tah1 to Pih1 allows for the formation of a stable complex. Based on our data, a model for an Hsp90-Tah1-Pih1 ternary complex is proposed.
Gómez-Mingot M, Alcaraz LA, MacIntyre DA, et al., 2011, Development of a novel analytical approach combining the quantification of amino acids, organic acids and glucose using HPLC-UV-Vis and HPLC-MS with screening via NMR, Analytical Methods, Vol: 4, Pages: 284-290
Gómez-Mingot M, Alcaraz LA, MacIntyre DA, et al., 2011, Development of a novel analytical approach combining the quantification of amino acids, organic acids and glucose using HPLC-UV-Vis and HPLC-MS with screening via NMR, Analytical Methods, ISSN: 1759-9660
A simple, rapid, sensitive and selective procedure based on the combination of HPLC-UV-Vis and HPLC-MS has been developed and single laboratory partially validated for the determination of a set of 13 analytes present in a commercially available IVF medium utilising small sample volumes (20–30 μL). The composition fingerprint of the complex sample obtained by NMR spectroscopy in 11 minutes provided identification based on a screening of the metabolomic profile. HPLC-MS allowed the glucose–sodium adduct to be measured accurately and the working and linear ranges achieved were 0.028–0.389 mmol L−1 with a detection limit of 13 μM. HPLC-UV-Vis allowed accurate concentrations of pyruvic and lactic acids with linear ranges over 0.005–0.1 mmol L−1 with a limit of detection of 28 μM for pyruvic acid to be determined in 8 minutes, while lactic acid presented a linear range over 0.1–2 mmol L−1 with a limit of detection of 1.2 mM possible. The use of HPLC-UV-Vis allowed the chromatographic separation of 8 amino acids (aspartate, glutamate, serine, glycine, asparagine, glutamine, alanine, and proline), the dipeptide alanyl-glutamine and taurine previous to a chemical derivatization, providing a total run time of 40 minutes. The method was partially validated to show a linear range of 0.028–0.280 mmol L−1 with detection limits ranging between 1 and 30 μM. Development of the analytical approach provided determination and quantification of a set of 13 analytes from a very complex sample. Although well established analytical techniques were used here, combinatory methodologies were partially validated for the first time to this purpose. The novelty of the combination of techniques relies on a screening tool and a strategy to the future evaluation and an improved assessment of human embryo viability.
MacIntyre DA, Melguizo Sanchis D, Jimenez B, et al., 2011, Characterisation of Human Embryonic Stem Cells Conditioning Media by <SUP>1</SUP>H-Nuclear Magnetic Resonance Spectroscopy, PLOS ONE, Vol: 6, ISSN: 1932-6203
- Author Web Link
- Open Access Link
- Cite
- Citations: 18
Jimenez B, Montoliu C, MacIntyre DA, et al., 2010, Serum metabolic signature of minimal hepatic encephalopathy by (1)H-nuclear magnetic resonance, J. Proteome Res., Vol: 9, Pages: 5180-5187, ISSN: 1535-3907
MacIntyre DA, Jimenez B, Lewintre EJ, et al., 2010, Serum metabolome analysis by 1H-NMR reveals differences between chronic lymphocytic leukaemia molecular subgroups, Leukemia, Vol: 24, Pages: 788-797, ISSN: 1476-5551
Mori M, Jiménez B, Piccioli M, et al., 2008, The solution structure of the monomeric copper, zinc superoxide dismutase from Salmonella enterica: structural insights to understand the evolution toward the dimeric structure., Biochemistry, Vol: 47, Pages: 12954-12963
The structure of the SodCII-encoded monomeric Cu, Zn superoxide dismutase from Salmonella enterica has been solved by NMR spectroscopy. This represents the first solution structure of a natural and fully active monomeric superoxide dismutase in solution, providing information useful for the interpretation of the evolutional development of these enzymes. The protein scaffold consists of the characteristic beta-barrel common to the whole enzyme family. The general shape of the protein is quite similar to that of Escherichia coli Cu, Zn superoxide dismutase, although some differences are observed mainly in the active site. SodCII presents a more rigid conformation with respect to the engineered monomeric mutants of the human Cu, Zn superoxide dismutase, even though significant disorder is still present in the loops shaping the active site. The analysis of both dynamics and hydration properties of the protein in solution highlights the factors required to maintain the fully active and, at the same time, monomeric protein. This study provides novel insights into the functional differences between monomeric and dimeric bacterial Cu, Zn superoxide dismutases, in turn helping to explain the convergent evolution toward a dimeric structure in prokaryotic and eukaryotic enzymes of this class.
Bertini I, Jiménez B, Pierattelli R, et al., 2008, Protonless 13C direct detection NMR: characterization of the 37 kDa trimeric protein CutA1., Proteins, Vol: 70, Pages: 1196-1205
The major limitation of nuclear magnetic resonance spectroscopy arises from the increase of nuclear transverse relaxation rates with increasing molecular mass. This causes reduction in spectral resolution and coherence transfer efficiency. The use of 2H-labeling to eliminate 1H-mediated relaxation pathways and the constructive use of cross correlation effects (TROSY, CRINEPT) alleviate the phenomenon. An alternative approach is to use direct detection of heteronuclei. Specifically designed 13C direct detection experiments can complement the set of 1H-based NMR experiments commonly used for structure determination providing an additional source of information less affected by the detrimental transverse relaxation effect. We applied this novel methodology to the study of the CutA1 protein (12.3 kDa) from E. coli that forms a homotrimer in solution with a total molecular mass of 37 kDa. In this work we demonstrate that the information available from 13C direct detection experiments makes it possible to completely assign the NMR resonances of the backbone of this 37 kDa trimeric protein without the need of deuteration. The structural and dynamical knowledge obtained for this system may contribute to understand its biological role.
Jiménez B, Mori M, Battistoni A, et al., 2007, NMR assignment of reduced form of copper, zinc superoxide dismutase from Salmonella enterica., Biomol NMR Assign, Vol: 1, Pages: 65-67
Almost complete assignment (97%) of NMR resonances was obtained for the reduced, Cu(I), form of prokaryotic CuZnSOD from Salmonella enterica. 13C direct detection was used to complement the standard bouquet of 1H detected triple resonance experiments and contributed to the identification of proline backbone resonances and to side chains assignments of Asx, Glx and aromatic rings. This is the only complete assignment available for monomer SOD from prokaryotic organisms.
Balayssac S, Jiménez B, Piccioli M, 2006, 13C direct detected COCO-TOCSY: a tool for sequence specific assignment and structure determination in protonless NMR experiments., J Magn Reson, Vol: 182, Pages: 325-329, ISSN: 1090-7807
A novel experiment is proposed to provide inter-residue sequential correlations among carbonyl spins in (13)C detected, protonless NMR experiments. The COCO-TOCSY experiment connects, in proteins, two carbonyls separated from each other by three, four or even five bonds. The quantitative analysis provides structural information on backbone dihedral angles phi as well as on the side chain dihedral angles of Asx and Glx residues. This is the first dihedral angle constraint that can be obtained via a protonless approach. About 75% of backbone carbonyls in Calbindin D(9K), a 75 amino acid dicalcium protein, could be sequentially connected via a COCO-TOCSY spectrum. 49(3)J(C')(C') values were measured and related to backbone phi angles. Structural information can be extended to the side chain orientation of aminoacids containing carbonyl groups. Additionally, long range homonuclear coupling constants, (4)J(CC) and (5)J(CC), could be measured. This constitutes an unprecedented case for proteins of medium and small size.
Balayssac S, Jiménez B, Piccioli M, 2006, Assignment strategy for fast relaxing signals: complete aminoacid identification in thulium substituted calbindin D 9K., J Biomol NMR, Vol: 34, Pages: 63-73, ISSN: 0925-2738
Paramagnetic proteins generally contain regions with diverse relaxation properties. Nuclei in regions far from the metal center may behave like those in diamagnetic proteins, but those closer to the metal experience rapid relaxation with accompanying line broadening. We have used a set of NMR experiments optimized to capture data from these various concentric regions in assigning the signals from a paramagnetic Calbindin D 9K derivative in which one of the two calcium ions has been replaced by thulium(III). Normal double- and triple-resonance experiments with 1H detection were used in collecting data from nuclei in the diamagnetic-like region; these approaches identified signals from fewer than 50% of the amino acid residues (those with d > 17.5 A from thulium(III)). Paramagnetism-optimized two-dimensional NMR experiments with 1H detection were used in collecting data from nuclei in the next nearer region (d > 15 A). Standard (d > 14 A) and optimized (d > 9 A) 13C direct-detection experiments were used to capture data from nuclei in the next layer. Finally nuclei closest to the metal were detected by one-dimensional 13C (d > 5 A) and one-dimensional 15N data collection (d > 4.2 A). NMR signals were assigned on the basis of through-bond correlations and, for signals closest to the metal, pseudocontact shifts. The latter were determined from chemical shift differences between assigned signals in thulium(III) and lanthanum(III) derivatives of Calbindin D 9K and they were interpreted on the basis of a structural model for the lanthanide-substituted protein. This approach yielded assignments of at least one resonance per amino acid residue, including those in the thulium(III) coordination sphere.
Bertini I, Jiménez B, Piccioli M, et al., 2005, Asymmetry in 13C-13C COSY spectra provides information on ligand geometry in paramagnetic proteins., J Am Chem Soc, Vol: 127, Pages: 12216-12217, ISSN: 0002-7863
The relative intensity of Calpha-C' cross-peaks in homonuclear 13C COSY spectra depends on the relaxation properties of Calpha and C' spins, which, in the proximity of a paramagnetic center, are related to the metal-to-carbon distance. Their quantitative analysis has lead, for the cerium-substituted dicalcium protein, calbindin D9k, to the straightforward identification of peaks arising from metal-coordinating groups. The monodentate or bidentate metal binding mode of carboxylates was identified directly via NMR.
Alcaraz LA, Jiménez B, Moratal JM, et al., 2005, An NMR view of the unfolding process of rusticyanin: Structural elements that maintain the architecture of a beta-barrel metalloprotein., Protein Sci, Vol: 14, Pages: 1710-1722, ISSN: 0961-8368
The unfolding process of the blue copper protein rusticyanin (Rc) as well as its dynamic and D(2)O/H(2)O exchange properties in an incipient unfolded state have been studied by heteronuclear NMR spectroscopy. Titrations of apo, Cu(I), and Cu(II)Rc with guanidinium chloride (GdmCl) show that the copper ion stabilizes the folded species and remains bound in the completely unfolded state. The oxidized state of the copper ion is more efficient than the reduced form in this respect. The long loop of Rc (where the first ligand of the copper ion is located) is one of the most mobile domains of the protein. This region has no defined secondary structure elements and is prone to exchange its amide protons. In contrast, the last loop (including a short alpha-helix) and the last beta-strand (where the other three ligands of the metal ion are located) form the most rigid domain of the protein. The results taken as a whole suggest that the first ligand detaches from the metal ion when the protein unfolds, while the other three ligands remain bound to it. The implications of these findings for the biological folding process of Rc are also discussed.
Bertini I, Jiménez B, Piccioli M, 2005, 13C direct detected experiments: optimization for paramagnetic signals., J Magn Reson, Vol: 174, Pages: 125-132, ISSN: 1090-7807
To optimize 13C direct detected experiments for the observation of signals close to a paramagnetic center, we have assessed the sensitivity of different sequences based on CO-Cali coherence transfer. Features of CACO experiments were tested for Calbindin D9k, in which one of the two native Ca2+ ions is replaced by the paramagnetic Ce3+ ion. We have studied the comparison of single vs multiple quantum coherence transfer evolution as well as the influence of in-phase vs anti-phase detection of 13CO signals and finally the comparison of a coherence transfer step based on a CyO in plane with respect to a Cy ali in plane. The acquisition of the anti-phase component of the signal, accomplished by the removal of the last refocusing steps, allowed the identification of some signals unobserved with other pathways. The structural dependency of paramagnetism-induced nuclear relaxation is such that the identification of the most suitable coherence transfer pathway is not known "a priori" but it is driven by the relative proximity of Cali and CO to the paramagnetic center.
Jiménez B, Moratal JM, Piccioli M, et al., 2004, Methods in proteome and protein analysis, Methods in proteome and protein analysis, Editors: Kamp, Calvete, Choli-Papadopoulou, Publisher: Springer Verlag, ISBN: 9783540202226
Mobility Studies in Proteins by 15N Nuclear Magnetic Resonance: Rusticyanin as an Example.
Jiménez B, Poggi L, Piccioli M, 2003, Monitoring the early steps of unfolding of dicalcium and mono-Ce3+-substituted forms of P43M calbindin D9k., Biochemistry, Vol: 42, Pages: 13066-13073, ISSN: 0006-2960
Early steps of unfolding of P43M Calbindin D(9k) have been evaluated by NMR spectroscopy on the native dicalcium and on the paramagnetic monocerium-substituted derivative. Although at 2 M GdmHCl the protein core maintains its overall folding and structure, amide (15)N R(2) measurements and cross correlation rates between N-H dipole-dipole relaxation and (15)N CSA relaxation reveal a closer and stronger packing of the hydrophobic interactions in the protein as a response to the presence of denaturing agents in solution. A complete reorientation of the Met43 side chain toward the hydrophobic core is accomplished by the disappearance of the millisecond dynamics observed on the native form of Calbindin D(9k), while cross correlation rates provide evidence that the two-way hydrogen bond between Leu23 and Val61 is broken or substantially weakened. The substitution of the calcium ion in site II with the paramagnetic Ce(3+) ion allowed us to obtain a number of long-range nonconventional constraints, namely, pseudocontact shifts, which were used, together with the NOEs collected on the native state, to monitor subtle structural variations occurring in the non-native state of the protein. Although the average rmsd between the structures of native and non-native states is small (0.48 A), structural rearrangements could be reliably identified. Our results provide unprecedented information about the behavior of Calbindin D(9k) during the early steps of unfolding. Furthermore, they constitute strong evidence of the efficiency of paramagnetism-based constraints in monitoring subtle structural changes that are beyond the sensitivity of an approach based only on NOE.
Jiménez B, Piccioli M, Moratal J-M, et al., 2003, Backbone dynamics of rusticyanin: the high hydrophobicity and rigidity of this blue copper protein is responsible for its thermodynamic properties., Biochemistry, Vol: 42, Pages: 10396-10405, ISSN: 0006-2960
Local dynamics and solute-solvent exchange properties of rusticyanin (Rc) from Thiobacillus ferrooxidans have been studied by applying heteronuclear ((1)H, (15)N) NMR spectroscopy. (15)N relaxation parameters have been determined for the reduced protein, and a model-free analysis has been applied. The high average value of the generalized order parameter, S(2) (0.93), indicates that Rc is very rigid. The analysis of cross correlation rates recorded in both the reduced and the oxidized forms conclusively proves that Rc possesses the same dynamic features in both oxidation states. The accessibility of backbone amide protons to the solvent at different time scales has also been studied by applying specific heteronuclear pulse sequences and by H(2)O/D(2)O exchange experiments. These experiments reveal that rusticyanin is extremely hydrophobic. The first N-35 amino acids, not present in the other BCPs, protect the beta-barrel core from its interaction with the solvent, and thus, this is one of the main factors contributing to the hydrophobicity. Both characteristics (high rigidity and hydrophobicity) are maintained in the metal ion surroundings.
Donaire A, Jiménez B, Fernández CO, et al., 2002, Metal-ligand interplay in blue copper proteins studied by 1H NMR spectroscopy: Cu(II)-pseudoazurin and Cu(II)-rusticyanin., J Am Chem Soc, Vol: 124, Pages: 13698-13708, ISSN: 0002-7863
The blue copper proteins (BCPs), pseudoazurin from Achromobacter cycloclastes and rusticyanin from Thiobacillus ferrooxidans, have been investigated by (1)H NMR at a magnetic field of 18.8 T. Hyperfine shifts of the protons belonging to the coordinated ligands have been identified by exchange spectroscopy, including the indirect detection for those resonances that cannot be directly observed (the beta-CH(2) of the Cys ligand, and the NH amide hydrogen bonded to the S(gamma)(Cys) atom). These data reveal that the Cu(II)-Cys interaction in pseudoazurin and rusticyanin is weakened compared to that in classic blue sites (plastocyanin and azurin). This weakening is not induced by a stronger interaction with the axial ligand, as found in stellacyanin, but might be determined by the protein folding around the metal site. The average chemical shift of the beta-CH(2) Cys ligand in all BCPs can be correlated to geometric factors of the metal site (the Cu-S(gamma)(Cys) distance and the angle between the CuN(His)N(His) plane and the Cu-S(gamma)(Cys) vector). It is concluded that the degree of tetragonal distortion is not necessarily related to the strength of the Cu(II)-S(gamma)(Cys) bond. The copper-His interaction is similar in all BCPs, even for the solvent-exposed His ligand. It is proposed that the copper xy magnetic axes in blue sites are determined by subtle geometrical differences, particularly the orientation of the His ligands. Finally, the observed chemical shifts for beta-CH(2) Cys and Ser NH protons in rusticyanin suggest that a less negative charge at the sulfur atom could contribute to the high redox potential (680 mV) of this protein.
Bertini I, Donaire A, Jiménez B, et al., 2001, Paramagnetism-based versus classical constraints: an analysis of the solution structure of Ca Ln calbindin D9k., J Biomol NMR, Vol: 21, Pages: 85-98, ISSN: 0925-2738
The relative importance of paramagnetism-based constraints (i.e. pseudocontact shifts, residual dipolar couplings and nuclear relaxation enhancements) with respect to classical constraints in solution structure determinations of paramagnetic metalloproteins has been addressed. The protein selected for the study is a calcium binding protein, calbindin D9k, in which one of the two calcium ions is substituted with cerium(III). From 1823 NOEs, 191 dihedral angles, 15 hydrogen bonds, 769 pseudocontact shifts, 64 orientational constraints, 26 longitudinal relaxation rates, plus 969 pseudocontact shifts from other lanthanides, a final family with backbone r.m.s.d. from the average of 0.25 A was obtained. Then, several families of structures were generated either by removing subsets of paramagnetism-based constraints or by removing increasing numbers of NOEs. The results show the relative importance of the various paramagnetism-based constraints and their good complementarity with the diamagnetic ones. Although a resolved structure cannot be obtained with paramagnetism-based constraints only, it is shown that a reasonably well resolved backbone fold can be safely obtained by retaining as few as 29 randomly chosen long-range NOEs using the standard version of the program PSEUDYANA.
This data is extracted from the Web of Science and reproduced under a licence from Thomson Reuters. You may not copy or re-distribute this data in whole or in part without the written consent of the Science business of Thomson Reuters.