1. Identify the amino acids in this peptide 2. Which amino acid sidechains are protonated at physiological pH 3. What is the overall charge of this peptide H₂N O NH₂ IZ ZI OH IZ `ОН `ОН
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- QUESTION 2 Which peptide will most strongly attach to a carboxymethyl ion exchange column at pH 7.4? O N'-Glu-Lys-Arg-Asp-Tyr-C' O N'-Tyr-Arg-Lys-Arg-Gly-C' O N'-Gly-Glu-Gln-Asp-Asp-C' O N'-Val-Ala-Leu-lle-Arg-C' QUESTION 3 Which will be product of transcription from this double-stranded (ds) DNA sequence? 5' - atgttcaaagcctattcg - 3' 3' - tacaagtttcggataagc - 5' O YKFRIS O uacaaguuucggauaagc O auauucaaaaccuauuca Which will be product of transcription from this double-stranded (ds) DNA sequence? 5' - atgttcaaagcctattcg – 3' 3' - tacaagtttcggataagc – 5' O YKFRIS O uacaaguuucggauaagc O auguucaaagccuauucg Ο MFKAYS QUESTION 4 The structure of hemoglobin consists of a 2a and 2B heterotetramer. O quaternary O tertiary O secondary O primarypeptide Lys-Glu-Trp answer the following questions: Draw the structure of this peptide when all the ionizable groups are fully protonated. How many ionizable groups does this peptide have? Label the pKa for each of the ionizable groups in the structure above. Draw the appropriate titration curve for this peptide on graph paper starting at pH 0 and ending at pH 14. Label the x-axis, y-axis and the pKa Determine the overall net charge of this peptide at each full equivalent point (including 0) where the ionizable group is 100% deprotonated. Determine the pI for this peptide. Determine the average overall net charge of a mixture of this peptide in a solution where pH=8.5.Given this: Protein Isoelectric pH Molecular weight (kDa) Ovalbumin 4.6 45 Insulin 5.4 5.7 Fibrinogen 5.8 340 Y-globulin 6.6 160 Collagen 6.6 115 Hemoglobin (monomer) 7.1 16 Myoglobin 7.0 16.7 What is the appearance after visualization of a protein mixture containing the seven proteins above if isoelectric focusing is done?
- Examine the following peptide and the inserted table, and answer the 2 questions below: Thr-Glu-Pro-Ile-Val-Ala-Pro-Met-Glu-Tyr-Gly-Lys 1. Estimate the net charge of the peptide at pH 2.0. Explain. 2. Estimate the net charge of the peptide at pH 7.0. Explain.TABLE 3-1 Properties and Conventions Associated with the Amino Acids pk, values Amino acid Phenylalanine Tyrosine Tryptophan 5.66 5.48 9.59 5.89 Abbreviation/ symbol 6.14 Phe F Tyr Y Trp W pk, M, (-COOH) 165 181 204 . 1.83 2.20 2.38 Using the table 3-1, calculate the pl of Tyrosine. Hint: Consider the structure and overall charge of the molecule between each pka. pK₂2 (-NH) 9.13 9.11 9.39 PKR (R group) 10.07V-M-Y-F-E-N: This is the single-letter amino acid abbreviation for a peptide. What is the net charge of this peptide at pH 7.0? An enzyme called a protein tyrosine kinase can attach phosphates to the hydroxyl groups of tyrosine (Y). What is the net charge of the peptide at pH 7.0 after it has been phosphorylated by a tyrosine kinase? What is the likely source of phosphate used by the kinase for this reaction?
- Calculate the net charge of the following peptide at pH 2, pH 6 and pH 14 (use 2 and 9 as pKa values for the carboxylic acid and amino groups, respectively, use the attached table for the side chain pKa values). Ile-Leu-Ile-Lys-Glu-Cys-His-Glu-Met-Ile-Ser-Thr-Arg-TyrH CH₂ H₂C HC-CH3 CH₂ H H₂C (S) H₂C H CH₂ CH₂ CH₂ NH O C NH NH₂ a) Which of the following statements about this peptide are correct? Group of answer choices Treatment of this peptide with trypsin generates two products. This peptide is a substrate for carboxypeptidase A Treatment of this peptide with cyanogen bromide generates a pentapeptide and a tripeptide. Treatment of this peptide with chymotrypsin generates three products. Treatment of this peptide with elastase generates 2 products. None of the above statements are correct. b) What is the sequence of this peptide using one letter abbreviations? c) What is the pH which would correspond to the ionization of the peptide as drawn above? 1, 5, 7, 10, 14Question 11. // Hint: Isoelectric focusing separates proteins based on their pI values, and can separate proteins that only differ by a net charge of ±1.±1. Recall that an amino acid residue with a negatively charged R group has a relatively low isoelectric point (pI) where it has zero net charge. Likewise, an amino acid residue with a positively charged R group has a relatively high isoelectric point (pI) where it has zero net charge. Order from Low pH to High pH
- Peptide A: AFEQHSR Calculate the isoelectric point of Peptide A using the pKa values in Table 4-1 of Voet, Voet and Pratt. Assume that the pKa values for the N-terminal amino group and C- terminal carboxyl groups are 8.0 and 3.5, as previously discussed in class. 3.785 5.055 7.02 10.24 3.50*H3N -Č-H CH2 он Tyrosine a. Draw the structures of tyrosine as different pH values. Please refer to the lecture notes for the pKa values b. Identify the structures that exist at pH 2 c. Solve for the pl. The following diagram shows the biosynthesis of B12 coenzymes, starting with the vitamin. DMB is dimethylbenzimidazole. (a) What one additional substrate or cofactor is required by enzyme B? (b) Genetic deficiency in animals of enzyme C would result in exces- sive urinary excretion of what compound?