Thesis (Ph. D.)--Columbia University, 1944. / Vita. eContent provider-neutral record in process. Description based on print version record. Bibliography: p. 19-20.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1970. / Typescript. Vita. Description based on print version record. Includes bibliographical references.

Thesis (Ph. D.)--University of Chicago, 1939. / Lithoprinted. "Private edition, distributed by the University of Chicago libraries, Chicago, Illinois."

Thesis--Stockholm. / Extra t.p., with thesis statement, inserted. Bibliography: p. at end.

University, Diss., 2002--Heidelberg.

Anterior gradient-2 (AGR2) is an endoplasmic reticulum (ER)-resident protein that belongs to a member of protein disulphide isomerase (PDI) superfamily. AGR2 initially emerged as a dominant effector of basic biological properties in vertebrates such as specifying forebrain integrity and limb generation. Subsequent studies in mammals implicated the role of AGR2 as a pro-metastatic protein essential to cancer progression and drug resistance, asthma and inflammatory bowel disease. AGR2 protein is mainly overexpressed in a number of human cancers and involved in pathways for ER stress, protein folding, transcription regulation, and exosome formation. Hence, AGR2 protein represents a clinically-relevant oncoprotein in tumour emergence and survival. The aim of this thesis is to shed more light on the role of AGR2 in cancer development. AGR2 was previously shown that it could bind sequence-specifically to a linear peptide motif. In this study, hydrogen/deuterium exchange mass spectrometry was used to identify the dominant peptide-binding site on AGR2 by comparing the deuterium uptake between AGR2 and AGR2 with its ligand (linear peptide motif). The binding of the peptide was probed by making mutant series in the identified peptide ‘docking site’ region on AGR2. A consensus peptide-binding motif was then developed to identify potential cellular proteins that harbour this motif as potential AGR2 client proteins. Database mining using this consensus binding peptide demonstrated that transmembrane proteins were dominant class of proteins. Epithelial cell adhesion molecule (EpCAM), an oncogenic transmembrane protein, was chosen as putative AGR2 client proteins and their interaction was verified using both cell-free and cell-based assays. The AGR2 and EpCAM pathway dynamics were reconstituted and investigated in cells that do not endogenously express both proteins. Further, the expression of AGR2 and EpCAM were assessed in clinical tumour samples using immunohistochemistry. Proteomics screen using quantitative tandem mass tag (TMT) mass spectrometry on cells transiently overexpressing AGR2 were used to identify potential AGR2 client proteins and to find relevant dominant pathways affected by AGR2 signalling. Additionally, synthetic tools were devised to further dissect the function, regulation and ‘druggability’ of AGR2 protein. These tools include: i) isolation of high-affinity AGR2-binding synthetic antibodies from a phage-scFv (single-chain variable fragment) library; ii) engineering synthetic mini-protein (synPRO) containing copies of wild-type and mutated AGR2 linear peptide motif and; iii) engineering synthetic membrane protein model that bind to AGR2. In conclusion, the data presented hereby demonstrated a novel role of an ER-resident protein AGR2 which possess an intrinsic sequence-specific peptide binding for a subset of its client proteins and one function of this motif is to ensure proper maturation of client proteins to their final destination. Development of synthetic tools in this study can be further manipulated to disrupt AGR2 signalling and the fate of its binding proteins which in turn highlights a potentially ‘druggable’ stage in the oncogenic secretory pathway.

Made available in DSpace on 2014-10-09T12:25:00Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:02:30Z (GMT). No. of bitstreams: 1 01272.pdf: 1663870 bytes, checksum: 36585130fb56868e0560fa884ac0d06f (MD5) / Dissertacao (Mestrado) / IPEN/D / Escola Politecnica, Universidade de Sao Paulo - POLI/USP

Made available in DSpace on 2014-10-09T12:25:00Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:02:30Z (GMT). No. of bitstreams: 1 01272.pdf: 1663870 bytes, checksum: 36585130fb56868e0560fa884ac0d06f (MD5) / Dissertacao (Mestrado) / IPEN/D / Escola Politecnica, Universidade de Sao Paulo - POLI/USP

The longitudinal and transverse nuclear relaxation times, T₁ and T₂, have been measured in normal H₂ gas at 77.5°K in the pressure range 0.05 to 2 atmospheres. In this region T₁ goes through a minimum, and T₂ deviates significantly from a linear dependence on the density. Comparison of the experimental data with existing theory establishes the following results for the J=1 state of orthohydrogen: i. autocorrelation functions of the molecular angular momentum operators are exponential or nearly so, ii. the ratio of the correlation times , Ʈ₁, Ʈ₂, which are associated with operators of the form J₊, and J²₊ respectively, lies within the limits 0.6 ≤ Ʈ₁ / Ʈ₂ ≤ 1, iii. the splitting of the molecular Zeeman levels cannot be neglected as in the original Schwinger theory. T₁ for the proton and deuteron in HD gas and for the deuterons in normal D₂ gas was measured as a function of temperature and pressure in the range 20 to 373°K and 0 to 8 atmospheres. To within experimental error the dependence of T₁ on the density p is linear. In HD below 65°K, when only the J=0 and J=1 states of the molecule are appreciably populated, the temperature dependence of T₁/p is identical for both proton and deuteron, leading to a value of Ʈ₁/Ʈ₂ = 1,07/± 15% for the J=1 state of HD. Above 100°K, T₁/p for the proton is inversely proportional to the temperature, whereas for the deuteron T₁/p is almost temperature independent. The experimental results are interpreted as evidence that in HD gas the process of molecular reorientation is dominated by the anisotropic intermolecular force arising from the separation of the centres of mass and charge of the molecule. In D₂ gas two relaxation times were found, one associated with the S=1 spin state of paradeuterium and the other associated with the S=2 spin state of orthodeuterium. At 40°K (T₁/p)s=₂ appears to go through a minimum; the analogous quantity in H₂ measured by previous workers also goes through a minimum, but at 80°K. This is consistent with interpreting the minimum as a quantum mechanical diffraction effect. The J=2 component of (T₁/p)s=₂ however, does not go through a minimum, which suggests that the intermolecular interactions are significantly different for the J=1 and J=2 states of the molecule. / Science, Faculty of / Physics and Astronomy, Department of / Graduate

An experimental method for the detection of the d(D,ɣ)He⁴ reaction has been developed. It involves the use of a double focusing magnetic spectrometer in conjunction with a solid state counter mounted in the focal plane of the spectrometer, the counter determining both the energy and dE/dx of the incident particles from the reaction. The design and construction of a particle beam handling system to guide the particle beam from the Van de Graaff generator to the object point of the spectrometer has been completed and tested. Using this beam, the characteristics of the spectrometer and solid state counter have been determined and recorded. An attempt was made to detect the d(D,ɣ)He⁴ reaction but no directly useful information on the reaction crossection was obtained. However, utilizing the knowledge gained during this experiment, it should be possible to make a more exacting attempt at the reaction crossection determination. / Science, Faculty of / Physics and Astronomy, Department of / Graduate