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- More ratios. Through the use of nuclear magnetic resonance spectroscopy, it is possible to determine the ratio between the protonated and deprotonated forms of buffers. (a) Suppose the ratio of [ A- ]A I to [HA] is determined to be 0.1 for a buffer with pKar6.0.pKa = 6.0. What is the pH? (b) For a different buffer, 91974 suppose the ratio of [ A- ]lA J to [HA] is determined to be 0.1 and the pHpH is 7.0. In this case, what is the pKapKa of the buffer? (c) For another buffer with pKa=7.5PKa = 7.5 at pH 8.0pH 8.0, what is the expected ratio of [ A- ][A ] to [HA]? doIonization State of Histidine.Each ionizable group of an amino acid can exist in one of two states, charged or neutral. The electric charge on the functional group is determined by the relationship between its pKa and the pH of the solution. This relationship is described by the Henderson-Hasselbalch equation. 1.Histidine has three ionizable functional groups. Write the equilibrium equations for its three ion-izationsand assign the proper pKa for each ionization. Draw the structure of histidine in each ionization state.What is the net charge on the histidine molecule in each ionization state? 2.Which structure drawn in (1) corresponds to theionization state of histidine at pH 1, 4, 8, and12?Note that the ionization state can be approximated by treating each ionizable group independently. 3.What is the net charge of histidine at pH 1, 4, 8, and 12? For each pH, will histidine migrate to-ward the anode (+) or cathode (-) when placed in an electric field?Cyclic forms of D-glucose: Haworth formula The furanose and pyranose forms of D-glucose can be presented using Haworth formula. The following templates are used for this formula. Note the numbering of the carbon atoms and the anomeric carbon (this carbon defines the alpha and beta isomer). Furanose template (furan) Pyranose template (pyran) anomeric carbon anomeric carbon Compare the Fisher and Haworth formula of D-glucofuranose and D-glucopyranose. Notice that the groups attached to C-1, C-2, and C-3 on the left side of the Fisher formula are written above the plane of the furanose template. Fisher formula Haworth formula H FOH H-2 HOH,č 5 -OH FOH- H HO-3H 4 он H 1 H-4 H-5 H OH 3 6 CH2OH H OH Alpha-D-glucofuranose Alpha-D-glucofuranose HOH2C- он OH но H FOH- OH Но H H. OH H ČH2OH Beta-D-glucofuranose Beta-D-glucofuranose
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- Peptides. 1. Draw the peptide Ala-Glu-Gly-Lys, as it would occur at physiological pH = 7.4. The R groups of Ala and Gly are not acidic or basic, therefore do not have a pka and the charge on these R groups is therefore independent of pH. Glu is acidic and Lys is basic, therefore the charge on these amino acids is pH dependent. The pKas are shown below. pKa N-term = 9.0 C-term = 3.5 Glu4.1 Lys = 10.5 2. Draw a circle around the peptide bonds. 3. Label the C-terminus and the N-terminus. 4. What is the overall charge on the peptide at pH 7.4?Thin Layer chromatography (TLC). Explain the biochemical principle behind the separation of carbohydrates molecules by TLC as performed in practical 5. Explain which properties molecules must have to travel a short and large distance, respectively. Ketohexose sugars can form 8 different stereoisomers. How many of those isomers can be distinguished and resolved by TLC as performed in the practical and why? no more than 100 words total. Draw Haworth projections for the following: (a) CHO in a-furanose form. Name the sugar. H-C-OH | HO-C-H H-C-OH CH,OH (b) The L isomer of (a) (c) a-D-GlcNAc (d) a-D-Fructofuranose
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