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- TPCK and TLCK are irreversible inhibitors of serine proteases. One ofthese inhibits trypsin and the other chymotrypsin. Which is which? Explainyour reasoning. Suggest the effects of each of the following mutations on the physiologicalrole of chymotrypsinogen:(a) R15S(b) C1S(c) T147SWhat effect does binding of the IRF protein to the IRE in the mRNA encoding ferritin have on the production of ferritin? Briefly explain why this effect is observed.During SDS-PAGE, glycoproteins migrate as relatively diffuse bands, whereas nonglycosylated proteins typically migrate as narrow, well-defined bands. Explain the reason for this difference in electrophoretic behavior.
- The first 32 amino acids from the N terminus of the protein bovine angiogenin were determined by Edman degradation and have the sequence:AQDDYRYIHFLTQHYDAKPKGRNDEYCFNMMK(a) Identify the sites of cleavage during trypsin-catalyzed hydrolysis of this protein.(b) What are the cleavage sites using chymotrypsin?The intermediates A, B, C, D, E, and F all occur inthe same biochemical pathway. G is the product of thepathway, and mutants 1 through 7 are all G−, meaningthat they cannot produce substance G. The followingtable shows which intermediates will promote growthin each of the mutants. Arrange the intermediates inorder of their occurrence in the pathway, and indicatethe step in the pathway at which each mutant strain isblocked. A + in the table indicates that the strain willgrow if given that substance, an O means lack of growth.SupplementsMutant A B C D E F G1 + + + + + O +2 O O O O O O +3 O + + O + O +4 O + O O + O +5 + + + O + O +6 + + + + + + +7 O O O O + O +b. Compounds A, B, C, and D are known to be intermediates in the pathway for production of protein E. To determine where the block in protein-E production occurred in each individual, the various intermediates were given to each individuals cel Is in culture. After a few weeks of growth with the intermediate, the cells were assayed for the production of protein E. The results for each individuals cells are given in the following table. A plus sign means that protein E was produced after the cells were given the intermediate listed at the top of the column. A minus sign means that the cells still could not produce protein E even after being exposed to the intermediate at the top of the column. Denote the point in the pathway in which each individual is blocked.
- a. Compounds A, B, C, and D are known to be intermediates in the pathway for production of protein E. To determine where the block in protein-E production occurred in each individual, the various intermediates were given to each individuals cel Is in culture. After a few weeks of growth with the intermediate, the cells were assayed for the production of protein E. The results for each individuals cells are given in the following table. A plus sign means that protein E was produced after the cells were given the intermediate listed at the top of the column. A minus sign means that the cells still could not produce protein E even after being exposed to the intermediate at the top of the column. Draw the pathway leading to the production of protein E.How do you suppose the serine/threonine acetylase activity of YopJ might interfere with TAK1 activation?In the galactose operon of E. coli, a repressor, encoded by the galR gene, binds to an operator site,galo, to regulate the expression of three structuralgenes, galE, galT, and galK. Expression is inducedby the presence of galactose in the media. For eachof the strains listed, would the cell show constitutive, inducible, or no expression of each of the structural genes? (Assume that galR− is a loss-of-functionmutation.)a. galR−galo+galE+galT+galK+b. galR+galocgalE+galT+galK+ c. galR−galo+galE+galT+galK−/galR+galo+galE−galT+galK+d. galR−galocgalE+galT+galK−/galR+galo+galE−galT+galK+
- In the galactose operon of Escherichia coli, a repressor, encoded by the galR gene, binds to an operator site, galo, to regulate the expression of three structural genes, galE, galT, and galK. Expression is induced by the presence of galactose in the media. For each of the strains listed, would the cell show constitutive, inducible, or no expression of each of the structural genes? (Assume that galR−is a loss-of-function mutation.) galR− galo+ galE+ galT+ galK+ galR+ galoc galE+ galT+ galK+ galR− galo+ galE+ galT+ galK−/ galR+ galo+ galE− galT+ galK+ galR− galoc galE+ galT+ galK−/ galR+ galo+ galE− galT+ galK+. An interesting mutation in lacI results in repressorswith 110-fold increased binding to both operator andnonoperator DNA. These repressors display a “reverse”induction curve, allowing β-galactosidase synthesis inthe absence of an inducer (IPTG) but partly repressingβ-galactosidase expression in the presence of IPTG. Howcan you explain this? (Note that, when IPTG binds a repressor, it does not completely destroy operator affinity,but rather it reduces affinity 110-fold. Additionally, ascells divide and new operators are generated by thesynthesis of daughter strands, the repressor must findthe new operators by searching along the DNA, rapidlybinding to nonoperator sequences and dissociating fromthem.)If you were given a tube containing only a purified protein of interest but had unlimited time and resources, how would you determine the in vivo expression levels of that protein? Is there another level of regulation beyond the protein expression level which might not be observable with the method you described?
