Predict the outcome of the following hypothetical experiments and be sure to explain and be rational in addition to proposing the probable outcome of the experiment: -> you alter the Rab-GDI binding site so that it cannot interact with Rab-GDP
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Predict the outcome of the following hypothetical experiments and be sure to explain and be rational in addition to proposing the probable outcome of the experiment:
-> you alter the Rab-GDI binding site so that it cannot interact with Rab-GDP
-> you have identified a drug that can specifically deplete PI(4)P from the trans-Golgi of the cell. When you treat cells with this drug you hypothesize....
-> while investigating novel anti-cancer drugs you identify a compound that disrupts y-tubulin localization to the centrosome and disperses it throughout the cytosol. Why might this compound be a useful anti-cancer agent?
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- Which ONE of these statements is the most accurate definition of the mode of action of imatinib? Select one: A.It is a specific inhibitor of the BCR‐ABL1 fusion protein and blocks phosphatase activity by competing with adenosine triphosphate (ATP) binding B.It is a specific inhibitor of the BCR‐ABL1 fusion protein and blocks tyrosine kinase activity by competing with adenosine triphosphate (ATP) binding C.It is a specific inhibitor of the BCR‐ABL1 fusion protein and blocks tyrosine kinase activity by interaction with the enzyme site D.It is a specific inhibitor of the BCR‐ABL1 fusion protein and blocks phosphatase activity by interaction with the enzyme siteFrom experiments in which cells expressing normal myosin II heavy chain were altered to either lack (mhckA-) or overexpress (MHCKA ++) a myosin heavy chain kinase (MHCKA). For answering this question recall the earlier the variants on the myosin II heavy chain, in which three key threonines, normally subject to reversible phosphorylation, were altered in various ways: 3X Ser = Serines in place of Threonines 3X Ala = Alanines in place of Threonines 3X Asp = Aspartate in place of Threonines MHCKA is present at normal levels. Which of the two mutants (mhcp- or MHCP++) would be most likely to have a defect in cytokinesis?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?
- Measure the uptake of leucine by epithetial cells of the mouse intestine. Measurements of the rate of update of L-leucine, D-Leucine, and L-valine , with and without Na+ in the assay were perform and yield different results (see table below). A) What can you conclude about the properties and mechanism of leucine transporter? B) Would you expect L-leucine uptake to be inhibited by Ouabain, which is a cardiac glycoside drug treatment?You are studying the uptake of L-leucine by epithelial cells of the mouse intestine. Measurements of the rates of uptake of L-leucine and severalof its analogs, with and without Na+ in the assay buffer, yield the results given in the table below. What can you conclude about the properties and mechanism of the leucine transporter? Would you expect L-leucine uptake to be inhibited by ouabain?Briefly describe the following properties of the Rab and Arf GTPases: a) Size, structure and cellular localization (for structure I want to know if they are lipidated and any other unique features) , b) How are they activated and inactivated (i.e. include the GEFs and GAPs), c). Give an example of downstream cellular effects.
- Indicate (x) if the following statements about synthesis of proteins containing an ER signal sequence are True or False: True False i. Translation is initiated by ribosomes located on the ER membrane The signal recognition particle (SRP) binds a sequence of nonpolar (hydrophobic) amino acids. ii. iii. The ER signal sequence may be cleaved by signal peptidase on the cytoplasmic side of the ER membrane. iv. The part of a transmembrane protein that will ultimately be located outside of the cell is inserted into the lumen of the ER during translation. A stop transfer sequence is a series of polar amino acids that halts translocation of a newly synthesized peptide into the ER lumen.Treatment of cells with the drug brefeldin A (BFA) has the effect of uncoating Golgi membranes, resulting in a cell in which the vast majority of Golgi proteins are found in the ER. What inferences can be made from this observation regarding roles of coat proteins other than promoting vesicle formation? Predict what type of mutation in ARF might have the same effect as treating cells with BFA.The PYK gene codes for the expression of pyruvate kinase, which is one of the enzymestargeted for anti-cancer drug design. You have identified an RNAi that targets the mRNAof PYK gene. To study the effect of the RNAi towards pyruvate kinase, the respected RNAiis expressed in Saccharomyces cerevisiae. The level of pyruvate kinase can be detectedwith a fluorescent antibody.(a). Predict the result that you will obtain in recombinant S. cerevisiae that expresses therespected RNAi.(b). Compare the result in Q3a(i) with the wild-type S. cerevisiae.
- The protein AKT/PKB is a critical regulator of essential cellular processes, and dysregulation of AKT has been implicated in many diseases, including cancer. In the present study, we wanted to test if levels of protein AKT changed in in EGF-stimulated epithelial, human breast cancer cell line when treated with varying concentrations of a drug Chondramide (ChB; 30 nM or 100 nM) for different lengths of time (1 or 24 h). Cellular lysate was prepared for each experimental condition, and an equal concentration of protein was loaded into a single lane of the SDS-polyacrylamide gel for each sample. We used western blot to measure the total levels of AKT and the fraction of Akt that is phosphorylated on the serine residue at position 473 (p-AktS473) using a phosphorylation specific antibody. GAPDH was used a loading control. EGF 30 nM ChB 100 nM ChB P-Ak47 total Akt GAPDH 0 0 + 0 0 0 0 0 1h 24h 1h 24h 0 Why do you think the researchers included GAPDH in the figure? A. B. C. D. Control for…Drug X shown below is a kinase inhibitor used to treat multiple types of breast cancer. Breast cancer cells are treated with the drug and after 8 hr are lysed and the presence of various proteins is shown by a band on a western blot. The presence of the phosphorylated form of the protein is shown by the presence of a band when probed with antibodies that recognize the phosphorylated form of protein and are indicated by a p- in front of protein name (e.g. p-AKT). Based on this information, which kinase is likely the target and is inhibited by Justify your answer in 3-4 sentences. Drug X. F FF N. -N S IZ H₂N `N No Drug (+) Drug p-AKT AKT p-GSK3B GSK3B p-p70S6K p70S6KThe diagram below shows a closeup of regulatory proteins binding to one of the UASG elements near the GAL7, GALI0, and GALI genes, which code for the protein products needed for yeast to use the sugar galactose. The red triangle symbolizes an "effector" molecule that binds to Gal80p. In this hypothesis (which has since been shown to be incorrect), what could be happening to Gal80p when it is bound to the effector molecule that causes it to change its position and uncover the Gal4p transcriptional activation domain. Hint: think about what effector molecules do upon binding to proteins such as the the Lac repressor protein or the CAP protein. Galactose absent, glucose absent Gal80p. _Activation domain Gal4p dimer -Binding domain UASG Galactose present, glucose absent Activation domain Gal80p- Binding domain UASG For the toolbar, press ALT+F10 (PC) or ALT+FN+F10 (Mac).