Factor H (FH) is a key regulator of the complement system, the principal molecular component of innate immunity in humans. The tight regulation of the alternative pathway (AP) of complement by FH occurs on host cells as well as in fluid phase. FH regulation of AP is achieved through its C3b. Bb-decay accelerating activity and cofactor activity for C3b proteolysis by factor I. This study presents evidence that the first three CCP modules, i.e. FH~1-3, constitute the minimal unit with cofactor activity for factor I. The work presented in this thesis describes the recombinant protein expression and NMR-derived structure determination of two overlapping pairs, FH~1-2 and FH~2-3, together with the use of these structures to build a model of the FH~1-3 structure. A structural comparison with other C3bengaging proteins (namely factor B, complement receptor type 1 and decay accelerating factor) is presented and used to devise hypotheses as to the respective roles of the three modules during an encounter with the convertase. This thesis further describes an investigation of the structural effects of two disease-associated sequence variants in the context of FH~1-2: namely the single nucleotide polymorphism V62I linked to age-related macular degeneration, and the R53H mutation linked to atypical haemolytic uraemic syndrome.

The increased demand for electric vehicles in recent years has driven the development of Li ion battery technology, yielding interesting trends in cathode materials. The layered cathode material Li(Ni1/3Mn1/3Co1/3)O2 gives 30% more reversible lithium extraction than the earlier LiCoO2 and the “overlithiated” material Li(Li0.2Mn0.54Ni0.13Co0.13)O2 gives a semi-reversible capacity 25% higher than Li(Ni1/3Mn1/3Co1/3)O2.1,2 6,7Li MAS NMR and 7Li MATPASS NMR were used to investigate the relation between the lithium ion and metal positions within these materials. It was found that Li(Ni1/3Mn1/3Co1/3)O2 showed a preference for Li ions to associate with Co at high voltages, that Mn4+ and Ni2+ showed some association and that the metals were not highly ordered. Li(Li0.2Mn0.5Ni0.13Co0.13)O2 showed a decrease in transition metal layer lithium upon cycling, in agreement with previous models, an ordering of the metal ions with the reinsertion of the lithium ions and a significant change in structure on deep discharge.3 These results will hopefully lead to more accurate modelling of the materials, understanding of reversibility and to increased reversible capacities in future cathode materials. Additionally work was done to enable high rate in-situ NMR spectra in which spectra are obtained from a cell while cycling in the bore of an NMR spectrometer. A Teflon Swagelok-style cell was designed and the effectiveness of solenoid and saddle coils were tested. It was found that for a 6 mm diameter cathode with a Li metal anode, at least half of the signal intensity could be obtained with a saddle coil whereas the signal was not detected when using a solenoid coil. / Thesis / Master of Science (MSc)

abstract: Glycosaminoglycans (GAGs) are a class of complex biomolecules comprised of linear, sulfated polysaccharides whose presence on cell surfaces and in the extracellular matrix involve them in many physiological phenomena as well as in interactions with pathogenic microbes. Decorin binding protein A (DBPA), a Borrelia surface lipoprotein involved in the infectivity of Lyme disease, is responsible for binding GAGs found on decorin, a small proteoglycan present in the extracellular matrix. Different DBPA strains have notable sequence heterogeneity that results in varying levels of GAG-binding affinity. In this dissertation, the structures and GAG-binding mechanisms for three strains of DBPA (B31 and N40 DBPAs from B. burgdorferi and PBr DBPA from B. garinii) are studied to determine why each strain has a different affinity for GAGs. These three strains have similar topologies consisting of five α-helices held together by a hydrophobic core as well as two long flexible segments: a linker between helices one and two and a C-terminal tail. This structural arrangement facilitates the formation of a basic pocket below the flexible linker which is the primary GAG-binding epitope. However, this GAG-binding site can be occluded by the flexible linker, which makes the linker a negative regulator of GAG-binding. ITC and NMR titrations provide KD values that show PBr DBPA binds GAGs with higher affinity than B31 and N40 DBPAs, while N40 binds with the lowest affinity of the three. Work in this thesis demonstrates that much of the discrepancies seen in GAG affinities of the three DBPAs can be explained by the amino acid composition and conformation of the linker. Mutagenesis studies show that B31 DBPA overcomes the pocket obstruction with the BXBB motif in its linker while PBr DBPA has a retracted linker that exposes the basic pocket as well as a secondary GAG-binding site. N40 DBPA, however, does not have any evolutionary modifications to its structure to enhance GAG binding which explains its lower affinity for GAGs. GMSA and ELISA assays, along with NMR PRE experiments, confirm that structural changes in the linker do affect GAG-binding and, as a result, the linker is responsible for regulating GAG affinity. / Dissertation/Thesis / Doctoral Dissertation Biochemistry 2015

The quest for a complete understanding of mixtures is a challenge which has stimulated the development of several techniques. One of the most powerful NMR-based techniques is known as Diffusion-Ordered SpectroscopY (DOSY), in which it is possible to distinguish the NMR spectra of chemical species with different hydrodynamic radii, i.e. with different self-diffusion coefficients. It allows the study of intact mixtures, providing information on the interactions within the mixture and saving time and money compared to other techniques. Unfortunately, DOSY is not very effective when signals overlap and/or the diffusion coefficients are very similar. This drawback has led to the development of new methods to overcome this problem. The present investigation is focused on developing some of these. Most DOSY datasets show multiplet phase distortions caused by J-modulation. These distortions not only hinder the interpretation of spectra, but also increase the overlap between signals. The addition of a 45º purging pulse immediately before the onset of acquisition is proposed as a way to remove the unwanted distortions. Most DOSY experiments use 1H detection, because of the higher sensitivity which is generally achieved. However, acquiring spectra with other nuclei such as 13C can reduce overlap problems. Two new sequences have been developed to maximize the sensitivity of heteronuclear DOSY experiments. In order to increase resolving power, it is also possible to incorporate another variable into diffusion experiments as a further dimension. If this results in an approximately trilinear dataset (each dimension varying independently), it is possible to extract physically meaningful information for each component using multivariate statistical methods. This is explored for the cases where the new variable is either the relaxation behaviour or the concentration variation (which can be measured during a reaction or in a set of samples with different concentrations for each component). PARAllel FACtor (PARAFAC) analysis can obtain the spectra, diffusional decay and relaxation evolution or kinetics for each of the components. In a completely different approach, the separation power of liquid chromatography has been combined in a novel way with the NMR potential for elucidating structures. NMR has been used previously as a precise way to measure average flow velocities, even in porous media. Using this capability to detect the different average velocities of solutes that occur in chromatographic columns ought to provide a new way of analysing mixtures with the same potential as LC-NMR, but faster and more simple. In such a flow system, a chromatographic column is introduced into the NMR probe and a 2D dataset is acquired and Fourier transformed to obtain the velocity distribution for each of the detected NMR signals.

We present nanoscale nuclear magnetic resonance (NMR) measurements performed with nitrogen-vacancy (NV) centers located down to about 2 nm from the diamond surface. NV centers were created by shallow ion implantation followed by a slow, nanometer-by-nanometer removal of diamond material using oxidative etching in air. The close proximity of NV centers to the surface yielded large 1H NMR signals of up to 3.4 lT-rms, corresponding to ~330 statistically polarized or ~10 fully polarized proton spins in a (1.8 nm)3 detection volume.

No description available.

Study of thermoresponsive porphyrins and their supramolecular complexes Abstract: We studied the water-soluble artificial compound meso-tetrakis{3,4,5-tris[2-(2-(2- methoxyethoxy)ethoxy)ethoxy]phenyl}porphyrin prepared at NIMS, Japan, using the high-resolution NMR spectroscopy experimental method. We observed its LCST-type phase separation and applied the Flory-Huggins theory of polymer solutions in order to find its phase diagram (binodal and spinodal curves of the phase separation) and we also obtained molar enthalpies, entropies and critical temperatures of its phase separation; from the Flory-Huggins theory we discovered that its molecules form dimers in aqueous solutions. We also studied its host-guest interactions with the S-camphorsulfonic acid; we learned that the porphyrin binds cations and the porphyrin dimers break down when dissolved cations are available for complexation. We observed no phase separation in chloroform. We obtained no proof of the existence of molecular stacks larger than dimers. 1

Certain types of porphyrins can be used as achiral agent for determination of enantiomeric excess (ee) of chiral molecules. Particular organic chiral molecule (guest) and porphyrin (host) form host-guest complex while inducing nonequiv- alency of particular proton resonances in symmetrical host. It causes splitting of NMR signals linearly dependent on ee of guest. In this work we investigated com- plexation of di-brombenzylated oxoporphyrin with chiral camphorsulfonic acid. NMR titration revealed that they form complex with 1:1 stoichiometry with as- sociation constant K ≈ 5 × 104 l/mol. We confirmed linear dependence of split- ting of host β-protons on ee of guest. Low temperature measurements revealed two conformations of host-guest complex with population around 0.7:0.3 (at −60 ◦ C). DFT quantum mechanical computations at BLYP/3-21G* level revealed also two conformations with population 0.79:0.21. NMR shifts were computed on this geometries with method GIAO/PBE1PBE/6-31G(2df,2pd) and compared to experimental values. 1

Höchste Magnetfelder haben sich zu einem unverzichtbaren Werkzeug der Festkörperphysik entwickelt. Sie werden insbesondere verwendet, um die elektronischen Eigenschaften von modernen Materialien zu erforschen. Da Magnetfelder oberhalb von 45 Tesla nicht mehr mit statischen (z.B. supraleitenden) Feldern zu erreichen sind, haben sich weltweit verschiedene Labore auf die Erzeugung gepulster Magnetfelder mit angestrebten Maximalwerten von 100 Tesla spezialisiert. In der vorliegenden Arbeit werden Anwendungsmöglichkeiten der kernmagnetischen Resonanz (NMR) in gepulsten Magnetfeldern aufgezeigt. Es ist gelungen, die starke Zeitabhängigkeit der gepulsten Magnetfelder mittels NMR präzise zu vermessen. Die genaue Kenntnis des Magnetfelds nach dem Puls ermöglicht, die Zeitabhängigkeit aus den Daten zu entfernen, sodass auch eine kohärente Signal-Mittelung möglich ist. Davon ausgehend werden erstmalig Messungen der chemischen Verschiebung, der Knight Shift, der Spin-Gitter-Relaxationsrate 1/T1 und der Spin-Spin-Relaxationsrate 1/T2 diskutiert. Schließlich werden die im Zusammenhang mit gepulsten Magnetfeldern erarbeiteten Gleichungen in vereinfachter Form zur genauen Messung und Analyse des freien Induktions-Zerfalls von 19F Kernspins in Calciumfluorid verwendet. Durch Messung des Zerfalls über sechs Größenordnungen wird eine genaue Analyse bezüglich einer neuartigen Theorie ermöglicht, welche den Zerfall basierend auf der Annahme mikroskopischen Chaos\'' erklärt. Diese Theorie hat das Potenzial, zu einem tieferen Verständnis von Quantenchaos in makroskopischen Vielteilchensystemen zu führen.

The thesis is devoted to the study of in-chain impurities in spin 1/2 antiferromagnetic Heisenberg chains (S=1/2 aHC's)---a model which accompanies the research on magnetism since the early days of quantum theory and which is one of the few integrable spin systems. With respect to impurities it is special insofar as an impurity perturbs the system strongly due to its topology: there is no way around the defect. To what extend the one-dimensional picture stays a good basis for the description of real materials even if the chains are disturbed by in-chain impurities is an interesting question which is addressed in this work. For this purpose, Cu Nuclear Magnetic Resonance (NMR) measurements on the cuprate spin chain compounds SrCuO2 and Sr2CuO3 intentionally doped with nickel (Ni), zinc (Zn) and palladium (Pd) are presented. These materials are well known to be among the best realizations of the S=1/2 aHC model and their large exchange coupling constants allow the investigation of the low-energy dynamics within experimentally easily feasible temperatures. NMR provides the unique ability to study the static and dynamic magnetic properties of the spin chains locally which is important since randomly placed impurities break the translational invariance. Because copper is the magnetically active ion in those materials and the copper nuclear spin is most directly coupled to its electron spin, the NMR measurements have been performed on the copper site. The measurements show in all cases that there are changes in the results of these measurements as compared to the pure compounds which indicate the opening of gaps in the excitation spectra of the spin chains and the emergence of oscillations of the local susceptibility close to the impurities. These experimental observations are compared to theoretical predictions to clarify if and to what extend the already proposed model for these doped systems---the finite spin chain---is suitable to predict the behavior of real materials. Thereby, each impurity shows peculiarities. While Zn and Pd are know to be spin 0 impurities, it is not clear if Ni carries spin 1. To shed some light on this issue is another scope of this work. For Zn impurities, there are indications that they avoid to occupy copper sites, other than in the layered cuprate compounds. Also this matter is considered.