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A KDEL sequence is a sequence of amino acids in a protein that binds a KDEL receptor, which helps recycle it back to the ER. Name two techniques a scientist could use in combination to experimentally demonstrate the function of a KDEL sequence. (To help you think about how to answer this: What would happen if the KDEL sequence were nonfunctional? How could you observe the result?)
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- Fluorescence-activated cell sorting (FACS) is a powerful technique for separating cells according to their content of particular molecules. For example, a fluorescence-labeled antibody specific for a cell-surface protein can be used to detect cells containing such a molecule. Suppose that you want to isolate cells that possess a receptor enabling them to detect bacterial degradation products. However, you do not yet have an antibody directed against this receptor. Which fluorescence-labeled molecule would you prepare to identify such cells?The following quote is from a TiBS review article (Y. Shiloh,(2006) TiBS vol. 31 No7 Pages 402-410). Please explain what information would have been derived from genetics experiments and what information would have been based on biochemistry experiments. Explain how using both genetic and biochemistry has allowed us to greatly enhance our understanding of this field. ATM belongs to a conserved family of proteins termed the ‘PI3K-like protein kinases’( PIKKs), most of which possess a serine/threonine kinase activity and all of which, as their name indicates, contain a domain with motifs that are typical of the lipid kinase phosphatidylinositol3- kinase(PI3K) [39]. At present, the mammalian members of this family include five protein kinases: ATM, ataxia-telangiectasia- and Rad3-related (ATR), hSMG-1, mTOR (also known as FRAP) and the catalytic subunit of the DNA-dependent protein kinase (DNA-PK). Another member of the PIKK family is TRRAP, a protein component of histone…Describes a method known as Western blotting that can be used to detect a polypeptide that is translated from a particular mRNA. In this method, a particular polypeptide or protein is detected by an antibody that specifically recognizes a segment of its amino acid sequence. After the antibody binds to the polypeptide within a gel, a secondary antibody (which is labeled) is used to visualize the polypeptide as a dark band.For example, an antibody that recognizes α-galactosidase A couldbe used to specifically detect the amount of α-galactosidase A proteinon a gel. The enzyme α-galactosidase A is defective in individuals with Fabry disease, which shows an X-linked recessive pattern of inheritance. Amy, Nan, and Pete are siblings, and Pete has Fabry disease. Aileen, Jason, and Jerry are brothers and sister, and Jerry has Fabry disease. Amy, Nan, and Pete are not related to Aileen, Jason, and Jerry. Amy, Nan, and Aileen are concerned that they could be carriers of a defective…
- A C-terminal KDEL tetrapeptide was identified by “sequence gazing” as a possiblesignal for keeping resident ER proteins in the ER. This idea is now described in yourtextbook as established fact. What types of experiments do you think were performedto confirm that KDEL serves as an ER localization signal?Metabolic syndrome is a clustering of various conditions including high blood pressure, high blood sugar, excess body fat, high serum triglycerides, and low high-density lipoproteins. Scientists studying metabolic syndrome cultured fibroblast cells from a metabolic syndrome patient and a healthy volunteer and challenged the cells to 100 nM insulin. Ten minutes after the stimulus was applied, they used a protein immunoblot to measure the amount of phosphorylated insulin receptor in the cultures. Use the passage to answer the question. Some patients with metabolic syndrome can continue to gain weight even if they consume fewer calories than other human adults. Based on this finding, which conclusion is MOST likely true? A. Patients with metabolic syndrome do not eliminate as many calories during excretion. B. Patients with metabolic syndrome do not expend as much energy during muscle contractions. C. Patients with metabolic syndrome have more efficient…Metabolic syndrome is a clustering of various conditions including high blood pressure, high blood sugar, excess body fat, high serum triglycerides, and low high-density lipoproteins. Scientists studying metabolic syndrome cultured fibroblast cells from a metabolic syndrome patient and a healthy volunteer and challenged the cells to 100 nM insulin. Ten minutes after the stimulus was applied, they used a protein immunoblot to measure the amount of phosphorylated insulin receptor in the cultures. Use the passage to answer the question. Insulin, in addition to regulating blood glucose, also promotes satiety. Ghrelin, on the other hand, is a peptide hormone that promotes hunger. Which statement BEST predicts the effect of ghrelin release on the insulin receptor? A. It increases insulin receptor expression. B. It decreases insulin receptor expression. C. It increases insulin receptor phosphorylation. D. It decreases insulin receptor phosphorylation.
- Metabolic syndrome is a clustering of various conditions including high blood pressure, high blood sugar, excess body fat, high serum triglycerides, and low high-density lipoproteins. Scientists studying metabolic syndrome cultured fibroblast cells from a metabolic syndrome patient and a healthy volunteer and challenged the cells to 100 nM insulin. Ten minutes after the stimulus was applied, they used a protein immunoblot to measure the amount of phosphorylated insulin receptor in the cultures. Use the passage to answer the question. A scientist hypothesizes that dysregulated insulin signaling is responsible for the development of excess body fat in metabolic syndrome patients. Based on the data, which of these, if true, would BEST support this hypothesis? A. Phosphorylated insulin receptor decreases the use of free energy in fibroblasts. B. Phosphorylated insulin receptor increases the use of free energy in fibroblasts. C. Phosphorylated insulin receptor…Metabolic syndrome is a clustering of various conditions including high blood pressure, high blood sugar, excess body fat, high serum triglycerides, and low high-density lipoproteins. Scientists studying metabolic syndrome cultured fibroblast cells from a metabolic syndrome patient and a healthy volunteer and challenged the cells to 100 nM insulin. Ten minutes after the stimulus was applied, they used a protein immunoblot to measure the amount of phosphorylated insulin receptor in the cultures. Use the passage to answer the question. The level of insulin in the blood is regulated by negative feedback. Which observation MOST supports this model? A. Insulin secretion increases metabolic syndrome symptoms. B. Insulin secretion decreases metabolic syndrome symptoms. C. Insulin receptor phosphorylation increases insulin secretion. D. Insulin receptor phosphorylation decreases insulin secretion.The above answer did not address the question in the sense that the answer did not indicate which protein involved in the pathways could be measured in the nucleus of cells and be confident in the results? I reiterate the question as follows: You want to do a single Western blot to measure changes in signaling activity, regardless of which pathway is being activated. For which protein involved in these pathways could you measure the levels in the nucleus of cells and be confident in your results? Why?
- BugRx is a new biotechnology company in Cambridge, Massachusetts, developing human monoclonal antibodies made in insect cells. The CEO, Dr. Bugsbunny, has convened a Board Meeting to discuss research proposals and budgets. The VP of Research & Development, Karen Hucklebug, gives an update on the Factor 801A program. Factor 801A is new signal transduction molecule that is important for the growth of ovarian cancer cells. She explains that the structure of the antibody binding to 810A has been finalized and production is on schedule in Luv Bug, Ireland. The CMO, Dr. David Sprayberry, reminds the Board that several pathologists have reported high levels of Factor 801A in ovarian cancer cells and it is a significant prognostic factor. The American Cancer Society estimates that 14,070 women died of ovarian cancer in the United States in 2018. The VP of Marketing, Andrew Flynn, asks about the current treatments for ovarian cancer. David explains that most ovarian cancers spread through…BugRx is a new biotechnology company in Cambridge, Massachusetts, developing human monoclonal antibodies made in insect cells. The CEO, Dr. Bugsbunny, has convened a Board Meeting to discuss research proposals and budgets. The VP of Research & Development, Karen Hucklebug, gives an update on the Factor 801A program. Factor 801A is new signal transduction molecule that is important for the growth of ovarian cancer cells. She explains that the structure of the antibody binding to 810A has been finalized and production is on schedule in Luv Bug, Ireland. The CMO, Dr. David Sprayberry, reminds the Board that several pathologists have reported high levels of Factor 801A in ovarian cancer cells and it is a significant prognostic factor. The American Cancer Society estimates that 14,070 women died of ovarian cancer in the United States in 2018. Give an example of a technique that can be used to measure the affinity of the antibody for Factor 801A? Explain how it works David adds that an…BugRx is a new biotechnology company in Cambridge, Massachusetts, developing human monoclonal antibodies made in insect cells. The CEO, Dr. Bugsbunny, has convened a Board Meeting to discuss research proposals and budgets. The VP of Research & Development, Karen Hucklebug, gives an update on the Factor 801A program. Factor 801A is new signal transduction molecule that is important for the growth of ovarian cancer cells. She explains that the structure of the antibody binding to 810A has been finalized and production is on schedule in Luv Bug, Ireland. The CMO, Dr. David Sprayberry, reminds the Board that several pathologists have reported high levels of Factor 801A in ovarian cancer cells and it is a significant prognostic factor. The American Cancer Society estimates that 14,070 women died of ovarian cancer in the United States in 2018 Propose a protocol (~100 words) to study the efficacy of the antibody in this mouse model.