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- A dimeric binding protein has a KD1 of 50 nM and a Hill coefficient of 1.4. What is the fraction of the protein that has ligand bound at 17 nM concentration of ligand?Why is glycine the only amino acid commonly observed in peptides inconformations with phi and psi values which are not restricted to the values in the Ramachandran plot below?Determine the buffer capacity of Alanine and Histidine at pH 2.0 and 6.0 respectively and discuss the results.
- Look up the pKa values for trimethylamine, dimethylamine, methylamine, and ammonia. Predict the order of elution of these compounds from a cation-exchange column eluted with a gradient of increasing pH, beginning at pH 7.Asparagine and glutamine residues in proteins are deamidated in an aqueous solution to yield aspartate and glutamate, respectively, and ammonium. Proteins undergoing deamidation of one or more of their Asn or Gln groups have been detected by isoelectric focusing. Why would isoelectric focusing be effective in separating deamidated from non-deamidated protein? Compare the relative pI values of the deamidated and non-deamidated proteins.Look up the pKa values for trimethylamine, dimethylamine, ethylamine, and ammonia. Predict the order of elution of these compounds from a cation-exchange column eluted with a gradient of increasing pH, beginning at pH 7.
- Show by a series of equations (with structures) the first stage of the Edman method applied to a peptide that has leucine as its N-terminal residue.At what distances do hydrogen bonding, hydrophobic interactions and charge transfer interactions act to dominate the interactions between particles when at the isoelectric point? The options are Short range or Long range.. An Analytical Chemist is analyzing a protein sample analysis using a light scattering technique. If the formulation of the subject sample has included surfactants or salt additives, which of the following would adequately apply? . The additives would impact the ionic strength of the sample B. The impact of additives influence can be nullified when using a Light Scattering technique. C. The effect of formulation additives would impact the protein sample analysis, and should be considered as a factor when analyzing the protein aggregation using light scattering technique.
- Write with the reasons why a separation will occur when a mixture of polypeptides with isoelectric points 4,3, 8,7 and 9,2 is applied to a column containing an anion-exchanging filler.Calculate the percentage ionization (i.e. [base]/[acid]x100) in these amino acid side chains at the pH values specified (show your work for one of these amino acid chains): - Aspartate (pKa=4.1) at pH 5.4 - Arginine (pKa=12.5) at pH 11.8 - Histidine (pKa=6.0) at pH 8.0 - Tyrosine (pKa=10.9) at pH 8.0The ESI mass spectrum of the protein transferrin gave a regular series of peaks, with the most intense peaks having the following m/z values.(i) Suggest a likely reason for the observation of these peaks in the ESI mass spectrum of the intact protein. (ii) Use the above data to calculate the molecular weight of transferrin. Show your working and state any assumptions made.
