Cholera is caused by a toxin produced by the bacterium Vibrio cholerae. Ultimately, the toxin leads to the activation of ion channels by Protein Kinase A (PKA), indirectly causing water to leave the cell (following ions out of the cell). This loss of electrolytes and water from intestinal cells into the intestinal lumen, produces the watery diarrhea characteristic of a V.cholera infection. Would you expect lower or higher levels of PKA in cells exposed to the cholera toxin? If you were to measure CAMP levels in these cells, what would you expect to find? Why? How might the cholera toxin modify G-proteins to result in these cAMP levels?
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- The S (spike) glycoprotein of coronaviruses can engage receptors by the N-terminal domain, the C-terminal domain, or both. The C-terminal domains of both human SARS coronaviruses bind ACE2 (angiotensin-converting enzyme 2), whereas that of MERS-CoV binds DDP4 (dipeptidyl peptidase 4). In contrast, in mouse hepatitis coronavirus (MHV) the N-terminal domain of S binds CEACAM (cell adhesion molecule 1). You have identified a novel coronavirus with an S protein that displays similarity to MHV at the N terminus and to SARS at the C terminus. What are the molecules most likely to function as receptors and how will you test this?You’ve made a 25 mM stock of colchicine (a tubulin inhibitor; dissolved in distilled water), which you’d like to use to study the effect of tubulin on phagocytosis in T.pyriformis. Based on published research articles that you’ve read, you’d like to preincubate T. pyriformis with colchicine at a final, effective concentration of 75 µM. Assuming you need 40 µL of the cell+colchicine mixture for your experiment, please explain how you can use your 25 mM solution of colchicine, T. pyriformis cells, and distilled water to create your cell+colchicine mixture.Cholera is an acute, diarrheal disease caused by infection of the intestine with the gram-negative, motile bacterium Vibrio cholerae. After colonizing the small intestine, V. cholerae secretes cholera toxin (CTX), a protein that ribosylates the Gsa subunit of a G protein, thereby inhibiting its GTPase activity. This causes the dysregulation of a cellular chloride ion channel, resulting in efflux of ions and water from the infected enterocytes into the intestinal lumen. Studies have implicated sodium bioenergetics in the pathogenesis of V. cholerae in humans. An electrochemical gradient of Na+ known as the "sodium motive force" provides V. cholerae with the energy for key functions. The gradient is predominantly established by the Nat-pumping NADH:quinone oxidoreductase (Na-NQR) enzyme. Na+-NQR is a multi-substrate enzyme comprised of six distinct subunits designated A-F (Table 1) that are encoded within the same operon. In a process similar to proton pumping by Complex I of the…
- A gut epithelial cell line that has been genetically engineered to express a common cystic-fibrosis-causing CFTR mutation is accidentally cross- contaminated with cholera bacteria from another experiment. Given what you know of CFTR's role in both disease states, what is a likely outcome? Cl- rushes out of the cells. O CI- rushes into the cells. O CI- levels are unchanged but Na+ is driven out of the cells. CI- levels in the cells are unaffected.α-Bungarotoxin is a powerful neurotoxin found in the venom of a poisonous snake (Bungarus multicinctus). It binds with high specificity to the acetylcholine receptor (AChR; an integral membrane protein) andprevents its ion channel from opening. This interaction was used to purify AChR from the electric organ of torpedo fish.(a) Outline a strategy for using α-bungarotoxin covalently bound to chromatography beads to purify the AChR protein. (b) Outline a strategy for the use of [125I]α-bungarotoxin to purify the AChR protein.When William H. was helping victims after a devastating earthquake in a region not prepared to swiftly set up adequate temporary shelter, he developed severe diarrhea. He was diagnosed as having cholera, a disease transmitted through unsanitary water supplies contaminated by fecal material from infected indiv iduals. The toxin produced by cholera bacteria causes Cl- channels in the lurninal membranes of the intestinal cells to stay open, thereby increasing the secretion of Cl- from the cells into the intestinal tract lumen. By what mechanisms would Na+ and water be secreted into the lumen in conjunction with Cl- secretion? How does this secretory response account for the severe diarrhea that is characteristic of cholera?
- You are studying growth factor GFA, which you know stimulates the proliferation of goblet cells in theintestine.Goblet cells are responsible for producing and secreting mucin, a mixture of glycosaminoglycans thatprotects the intestinal wall.Some patients suffering from inflammatory bowel disease (IBD) appear to have fewer goblet cells, thereforeless mucin and less protection from toxins and various other pro-inflammatory factors.These patients also have mutations in the gene encoding the GFA receptor (GFAR) in goblet cells,GFAR is a receptor tyrosine kinase (RTK) that autophosphorylates in response to GFA binding, thusbecoming active.QUESTION:explain what changes in GFAR could be caused by these IBD-associatedmutations and why.Recent evidence points towards a global increase in cases of multi drug and total drug resistant forms of Mycobacterium tuberculosis, suggesting that the mycobacterium is becoming resistant to the antibiotic regimen used to treat this infectious disease. New therapies are clearly needed, which may emerge from an improved understanding of the mechanisms by which M. tuberculosis avoids cellular clearance by lysosomes following engulfment in macrophage phagosomes. We now report that one of the common anti-tubercular drug targets, the so-called RND permease multi substrate pumps, is capable of secreting mycolic acids, a form of lipid enriched in these mycobacteria that make up part of the pathogens cell wall, as a way to inhibit phago- lysosome fusion and survive within the host cell. Our findings provide a mechanism that may explain how these cholesterol like mycolic acids may inhibit the lysosomal protein NPC1, a lysosomal cholesterol transporter and the only mammalian member of the RND…The small fragments of some complement proteins initiate a local inflammatory response. Recent studies using mouse models of pulmonary inflammation (a model for human asthma) have found that mice deficient in the C3a receptor have greatly reduced disease symptoms when challenged with inhaled preparations containing extracts of the fungal pathogen Aspergillus fumigatus. Specifically, the C3a receptor-deficient mice showed reduced influx of granulocytes and lymphocytes into the lung and reduced fluid in the lung after challenge. What is the explanation for these findings?
- AAA is a protein found in both the cytosol as well as the nucleus. In its active state, it helps to facilitate the movement of proteins into and out of the nucleus. Specifically, AAA-alpha and AAA-beta binds to receptors on the nucleus and allows for materials to enter or exit through the channels associated with the receptors. They also play a role in dropping off cargo that import and export receptors hold onto. AAA functions are controlled by two other proteins: BBB and CCC. BBB binds onto AAA, while CCC removes BBB from AAA when they are bound. As a result, there is a BBB and CCC concentration gradient that forms between the cytosol and nucleus. During prophase I, is there a concentration gradient for BBB and CCC between the cytosol and the nucleus? Why? Is this also the case during the other phases of meiosis or the cell cycle?AAA is a protein found in both the cytosol as well as the nucleus. In this active state, it helps to facilitate the movement of proteins into and out of the nucleus. Specifically, AAA-alpha and AAA-beta binds to receptors on the nucleus and allows for materials to enter or exit through the channels associated with the receptors. They also play a role in dropping off cargo that import and export receptors hold onto. AAA functions are cotrolled by two other proteins: BBB and CCC. BBB binds onto AAA, while CCC removes BBB from AAA when they are bound. As a result, there is a BBB and CCC concentration gradient that forms between the cytososl and nucleus. During prophase I, is there a concentration gradient for BBB and CCC between the cytosol and the nucleus? Why? is this also the case during the other phases of meiosis or the cell cycle?Poliovirus is highly cytopathic both in cultured cells and in animals. Inoculation of mice with poliovirus leads to a robust antibody and cellular response. Explain why mice infected with a poliovirus mutant that cannot cause cell death have poor antibody and cellular responses. What would be one strategy to restore robust antibody and cellular responses in mice infected with this mutant virus?