Project description:Proteomic investigation on the glycosylation substrates and proteome effects of altering neisserial OTases within the proteome of N. gonorrhoeae MS11
Project description:Within the Burkholderia genus O-linked protein glycosylation is now known to be highly conserved at the pathway and glycosylation substrate levels. While inhibition of glycosylation has been shown to be detrimental to virulence in B. cenocepacia, little is known about the role of glycosylation in Burkholderia glycoproteins. Within this study we have sought to improve our understanding of the breadth and dynamics of the B. cenocepacia O-glycoproteome to identify glycoproteins which require glycosylation for functionality. Assessing the glycoproteome across multiple common culturing media (LB, TSB, and artificial sputum medium to simulate cystic fibrosis sputum-like conditions) we demonstrate at least 141 glycoproteins are subjected to glycosylation within B. cenocepacia K56-2. Leveraging this insight, we quantitively assessed the glycoproteome of B. cenocepacia using Data-Independent Acquisition (DIA) across culturing media and growth phases revealing most B. cenocepacia glycoproteins are express under all conditions. Examination of how the absence of glycosylation impacts the glycoproteome reveals only a subset of the glycoproteome (BCAL1086, BCAL2974, BCAL0525, BCAM0505 and BCAL0127) appear impacted by the loss of glycosylation. Assessing the proteomic and phenotypic impacts of the loss of these glycoproteins compared to glycosylation null strains revealing the loss of BCAL0525, and to a lesser extend BCAL0127, mirror the proteomic effects observed in the absence of glycosylation. Finally, we demonstrate the loss of glycosylation within BCAL0525 at Serine-358 results in both loss of motility and proteomic impacts on flagellar apparatus consistent with the loss of apparatus stability. Combined this work demonstrates that O-linked glycosylation of BCAL0525 is functionally important within B. cenocepacia.
Project description:Fatty liver disease and insulin resistance are common comorbidities, highlighting the potential for the liver to secrete factors that influence glycemic control. We assessed the influence of low vs high glucose on liver secreted extracellular vesicle secretion and composition
Project description:Transcriptional profiling of Arabidopsis in response to infection with TMV in the initial infection stage (0.5, 4 and 6 hours post inoculation). Analyze the relative transcriptome change of TMV*CP.MP- and TMV*rep-inoculated samples. TMV*rep vs. TMV*CP.MP transfected cells. 2 biological replicates conducted at each time (0.5, 4 and 6 hours post inoculation)
Project description:Fewer than 50% of patients develop vascular and valvular calcification, implying differential pathogenesis. Tissue-entrapped extracellular vesicles (EVs) are found in mineralization but their contents and functions are unstudied. Tissue EVs were isolated from normal and diseased human carotid arteries and aortic valves by enzymatic digestion, serial (ultra)centrifugation and density-gradient separation. Mass spectrometry characterized the density-gradient separation vs. ultracentrifugation.