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- Mammalian cells growing in culture were labeled with [3H]-thymidineto estimate the rate of DNA synthesis. The thymidine administered had aspecific activity of 3000 cpm/pmol. At intervals, samples of culture weretaken and acidified to precipitate nucleic acids. The rate of incorporation ofisotope into DNA was 1500 cpm/106 cells /min. A portion of culture wastaken to determine the specific activity of the intracellular dTTP pool, whichwas found to be 600 cpm/pmol.(a) What fraction of the intracellular dTTP is synthesized from the exogenous precursor?(b) What is the rate of DNA synthesis, in molecules per minute per cell ofthymine nucleotides incorporated into DNA?(c) How could you determine the specific activity of the dTTP pool?Equilibrium constants are always dependent upon temperature. In determining equilibrium constants for biochemical reactions the equilibrium constant also depends upon ion concentration and proton concentration. Write down an equation in differential form that describe the dependence of an equilibrium constant K for a protein binding to DNA, where the equilibrium constant, K is a function of Temperature, [NaCI], and [H*], i.e. dK(T, [NaCl], [H*]).High salt concentrations tend to cause protein aggregation. Suggest a way to identify proteins normalexpressed in particular bacterial species that can retaintheir solubility despite high salt conditions.
- Using the pKa data in as shown and the Henderson-Hasselbalch equation,calculate the approximate net charge on each of the four common ribonucleoside 5′-monophosphates (rNMPs) at pH 3.8. If a mixture of these rNMPs was placed in an electrophoresis apparatus, as shown, draw four bands to predict the direction and relative migration rate of each.Suggest a reasonable strategy for the specific phosphorylation of the5’ –OH group of a nucleoside.Bovine ribonuclease folds with ΔH° = -280 kJ mol-1 and ΔS° = -0.79 kJ mol-1 K -1. Assume ΔH° and ΔS° are independent of temperature. What fraction of bovine ribonuclease is unfolded at 42 °C?
- Fibrinogen contains tyrosine- O - sulfate. Propose an activated form of sulfate that could react in vivo with the aromatic hydroxyl group of a tyrosine residue in a protein to form tyrosine- O -sulfate.Given the undecapeptide un sequenced Ala2, Arg, Glu, Lys, Met, Phe, Ser, Thr, Trp, Val. Edman Degradation identified Ala. It is further analyzed and degraded to four fragments using trypsin- catalyzed hydrolysis and yielded the following Ala – Glu – Arg ;Thr – Phe – Lys ; Lys ; and Met – Ser – Trp – Val . To further analyze the undecapeptide A.A Sequence, Chymotrypsin -catalyzed hydrolysis identified two fragments Ala – Arg – Glu – Phe – Thr and Lys – Met – Ser – Trp – Val and lastly treatment with cyanogen bromide identified two fragments: Ala – Arg – Glu – Lys – Met – Phe – Thr – Val; and Trp – Ser what is the name of the undecapeptide?Purification of a protein of unknown structure has been achieved. The natural protein has a molecular weight of 240,000, according to size-exclusion chromatography. Using a concentration of 6 M guanidine hydrochloride in the chromatography, a single peak can be identified as the molecular weight (MW) 60,000 of a protein. B-mercaptoethanol (BME) and guanidine hydrochloride (GHC) are used in tandem to produce proteins with mass masses of 34,000 and 26,000, respectively. The structure of this protein can be inferred from these facts.
- It is known that double stranded DNA is denatured at low pH. pKa values should allow the determination of whether this is due to perturbation of the hydrogen bonding in A-T and/or G-C base pairs. The table gives values for the pKas of different protonated groups in the nucleobases.Nucleobase Position & pKa A N1, 3.5 G N7, 1.6; N1, 9.2 C N3, 4.2 T N3, 9.7a) Draw the A-T and G-C base pairs. - Label the bases with the one-letter code. – - Number the atoms in the rings and label the atom that attaches to the sugar. - Mark the groups that interact in normal…Mutants of Neurospora crassa that lack carbamoyl phosphate synthetase I (CPS I) require arginine in the medium in order to grow, whereas mutants that lack carbamoyl-phosphate synthetase II (CPS II) require a pyrimidine, such as uracil. A priori, one would expect the active CPS II in the arginine mutants to provide sufficient carbamoyl phosphate for arginine synthesis, and the active CPS I in the pyrimidine mutants to "feed" the pyrimidine pathway. Explain these observations.Given the following diagram of how protein AWESOME1 binds to it's target DNA, describe the potential effects of each of the 5 mutations shown below. The wild-type sequence of a helix #1 is also shown in the blue box, and all the mutations are in helix #1 (see numbers for identifying particular residues). a helix #1 R(1)-V-I-L-Y-F-W-I-M-Y-F-S-H-Y-W-R(16) #1 Predict the consequence of the following mutations: 1) Arg(1) to Glu 2) Arg(1) to Ala 3) Phe(6) to lle 4) Trp(7) to Phe 5) Met(9) to Pro in
