Project description:HDX-MS data revealing long-range allostery that affects the catalytic activity of the serine protease, murine urokinase plasminogen activator. The allostery is mediated by the disordered region connecting the N-terminal EGF and kringle domains to the protease domain.
Project description:Genome-wide association studies in multiple sclerosis (MS) identified a polymorphism (rs6897932) located in the coding region of the alpha chain of the cytokine receptor interleukin 7 receptor (IL7R) as a component that increases susceptibility to develop the disease. This single nucleotide polymorphism (SNP) affects the splicing of the primary transcript leading to genotype-defined transcript ratios encoding either a full length membrane spanning form or a soluble receptor chain. Genotyping at the IL7R locus reveals that the region can be described by four haplotypes. Interestingly, only one out of three haplotypes harbouring the associated SNP is positively associated with MS whereas the other two do not show association. The minor allele containing haplotype shows a reduced susceptibility to develop MS. We hypothesized that additional functional or phenotypic differences exist between individuals homozygous for haplotypes shown to have either positive, negative, or neutral effect, on susceptibility to develop MS. Gene expression profiles of CD4+ T cells from MS individuals before and after stimulation with IL7 were recorded. Haplotype-specific gene signatures were found indicating small alterations in IL7/IL7R signal processing/sensitivity through JAK/STAT and p38/MAPK14. We can not exclude that the obtained signatures result from differences within the CD4+ T cell compartment that, in fact, should be seen as a consequence of systemic haplotype-specific processing of homeostatic and proliferation signals transmitted through IL7/IL7R.
Project description:We have performed in solution hydrogen exchange (HDX) on peptide standards and bovine hemoglobin with subsequent quenching, pepsin digestion, and cold capillary electrophoretic separation coupled with mass spectrometry (MS) detection for the benchmarking of a lab built HDX-MS platform. Though neutral coated capillaries that eliminate electroosmotic flow provide fast separations with peak capaci-ties surpassing 200, uncoated capillaries with high electroosmotic flow achieve half as much peak ca-pacity with even faster separations but 30% higher deuterium retention for angiotensin II peptide standard. The data obtained from two different separation conditions on peptic digests of bovine he-moglobin shows strong agreement for relative deuterium uptake between methods and provides data amenable to software such as MS-Studio. Processed data from denatured hemoglobin vs the longest time point in this study (50,000s) also shows agreement with subunit interaction sites determined by crystallographic methods.
Project description:Genome-wide association studies in multiple sclerosis (MS) identified a polymorphism (rs6897932) located in the coding region of the alpha chain of the cytokine receptor interleukin 7 receptor (IL7R) as a component that increases susceptibility to develop the disease. This single nucleotide polymorphism (SNP) affects the splicing of the primary transcript leading to genotype-defined transcript ratios encoding either a full length membrane spanning form or a soluble receptor chain. Genotyping at the IL7R locus reveals that the region can be described by four haplotypes. Interestingly, only one out of three haplotypes harbouring the associated SNP is positively associated with MS whereas the other two do not show association. The minor allele containing haplotype shows a reduced susceptibility to develop MS. We hypothesized that additional functional or phenotypic differences exist between individuals homozygous for haplotypes shown to have either positive, negative, or neutral effect, on susceptibility to develop MS. Gene expression profiles of CD4+ T cells from MS individuals before and after stimulation with IL7 were recorded. Haplotype-specific gene signatures were found indicating small alterations in IL7/IL7R signal processing/sensitivity through JAK/STAT and p38/MAPK14. We can not exclude that the obtained signatures result from differences within the CD4+ T cell compartment that, in fact, should be seen as a consequence of systemic haplotype-specific processing of homeostatic and proliferation signals transmitted through IL7/IL7R. Samples of CD4+ cells were obtained from 7 MS patients (homozygous for Hap1 (3), Hap2 (2), Hap3 (2)). CD4+ cells were collected from peripheral blood, frozen and stored in liquid nitrogen. All samples were thawed and CD4+ cells were purified by magnetic bead separation. Purity and viability of cells was analyzed by Fluorescence Activated Cell Sorter (FACS). Total cellular RNA were extracted with TRIzol reagent and analyzed with the Human Gene 1.0 ST Array (affymetrix). IL7R haplotypes and susceptibility to develop MS: Hap1 homozygous <-> Risk <-> positive effect on MS susceptibility Hap2 homozygous <-> Hap2 <-> neutral effect on MS susceptibility Hap3 homozygous <-> Prot <-> neutral effect on MS susceptibility [Note: Haplotype nomenclature subject to revision.]
Project description:HDXMS raw data for various states of EFSAM including various mutants, and the apo and calcium-bound forms. The ID runs that were used to identify peptides in PLGS are also included.
Project description:We performed a retrospective study on CSF from 20 DMT-naïve MS patients to investigate the correlation between intrathecal immune proteins and clinical MS phenotype.
Project description:Comparison of two solid-phase extraction (SPE) methods for the identification and quantification of porcine retinal protein markers by LC MS/MS