9 research outputs found
The influence of nitrogen-15 proton-driven spin diffusion on the measurement of nitrogen-15 longitudinal relaxation times
The effect of nitrogen-15 proton-driven spin diffusion on quantitative N-15 T-1 measurements in solid proteins is investigated, and the impact on the measurement of dynamic parameters is assessed. A simple model of exchange between neighboring nitrogens is used to reproduce the evolution of N-15 spin systems whose longitudinal relaxation rates and exchange rates are compatible with experimental measurements. We show that the induced error in the measured T-1 and its effect on the determination of dynamics parameters is likely to be less than the current experimental error. The use of deuterated protein samples is shown to have a small but sometimes visible effect, and may also considerably slow down or even suppress the exchange of magnetization due to spin diffusion. (c) 2006 Elsevier Inc. All rights reserved
A project report about the new integrated seminar “broken bone” at the department of anatomy and cellbiology of the University of Ulm
Coherence transfer selectivity in two-dimensional solid-state NMR
In solid-state NMR, recoupling sequences are often used as the mixing elements of two-dimensional pulse sequences. We show that in many cases, the commutation properties of the mixing Hamiltonian leads to selectivity in the allowed coherence transfer processes. This property may lead to enhanced resolution in the cross-peaks of two-dimensional solid-state NMR spectra
Solid-state NMR spectroscopy of a paramagnetic protein: Assignment and study of human dimeric oxidized Cu-II-Zn-II superoxide dismutase (SOD)
[(C5H5)Ni{P(S)R2}2Ni(C5H5)], zweikernige Spin-Crossover-Komplexe: Synthese, Elektronenstruktur sowie Kinetik und Mechanismus der Isomerisierung
Observation of heteronuclear overhauser effects confirms the N-15-H-1 dipolar relaxation mechanism in a crystalline protein
Comparison of the integrin α4β7 expression pattern of memory T cell subsets in HIV infection and ulcerative colitis.
Anti-α4β7 therapy with vedolizumab (VDZ) has been suggested as possible immune intervention in HIV. Relatively little is known about the α4β7-integrin (α4β7) expression of different T-cell subsets in different anatomical compartments of healthy individuals, patients with HIV or inflammatory bowel disease (IBD). Surface expression of α4β7 as well as the frequency of activation, homing and exhaustion markers of T cells were assessed by multicolour flow cytometry in healthy volunteers (n = 15) compared to HIV infected patients (n = 52) or patients diagnosed with ulcerative colitis (UC) (n = 14), 6 of whom treated with vedolizumab. In addition, lymph nodal cells (n = 6), gut-derived cells of healthy volunteers (n = 5) and patients with UC (n = 6) were analysed. Additionally, we studied longitudinal PBMC samples of an HIV patient who was treated with vedolizumab for concomitant UC. Overall, only minor variations of the frequency of α4β7 on total CD4+ T cells were detectable regardless of the disease status or (VDZ) treatment status in peripheral blood and the studied tissues. Peripheral α4β7+ CD4+ T cells of healthy individuals and patients with UC showed a higher activation status and were more frequently CCR5+ than their α4β7- counterparts. Also, the frequency of α4β7+ cells was significantly lower in peripheral blood CD4+ effector memory T cells of HIV-infected compared to healthy individuals and this reduced frequency did not recover in HIV patients on ART. Conversely, the frequency of peripheral blood naïve α4β7+ CD4+ T cells was significantly reduced under VDZ treatment. The results of the current study will contribute to the understanding of the dynamics of α4β7 expression pattern on T cells in HIV and UC and will be useful for future studies investigating VDZ as possible HIV cure strategy
Overall Structural Model of NS5A Protein from Hepatitis C Virus and Modulation by Mutations Confering Resistance of Virus Replication to Cyclosporin A
Hepatitis
C virus (HCV) nonstructural protein 5A (NS5A) is a RNA-binding
phosphoprotein composed of a N-terminal membrane anchor (AH), a structured
domain 1 (D1), and two intrinsically disordered domains (D2 and D3).
The knowledge of the functional architecture of this multifunctional
protein remains limited. We report here that NS5A-D1D2D3 produced
in a wheat germ cell-free system is obtained under a highly phosphorylated
state. Its NMR analysis revealed that these phosphorylations do not
change the disordered nature of D2 and D3 domains but increase the
number of conformers due to partial phosphorylations. By combining
NMR and small angle X-ray scattering, we performed a comparative structural
characterization of unphosphorylated recombinant D2 domains of JFH1
(genotype 2a) and the Con1 (genotype 1b) strains produced in <i>Escherichia coli</i>. These analyses highlighted a higher intrinsic
folding of the latter, revealing the variability of intrinsic conformations
in HCV genotypes. We also investigated the effect of D2 mutations
conferring resistance of HCV replication to cyclophilin A (CypA) inhibitors
on the structure of the recombinant D2 Con1 mutants and their binding
to CypA. Although resistance mutations D320E and R318W could induce
some local and/or global folding perturbation, which could thus affect
the kinetics of conformer interconversions, they do not significantly
affect the kinetics of CypA/D2 interaction measured by surface plasmon
resonance (SPR). The combination of all our data led us to build a
model of the overall structure of NS5A, which provides a useful template
for further investigations of the structural and functional features
of this enigmatic protein