1. You are trying to purify a protein that has a total ratio of hydrophobic to h
ID: 167820 • Letter: 1
Question
1. You are trying to purify a protein that has a total ratio of hydrophobic to hydrophilic amino acids of 70:30 so you decide to purify the Lysate using HIC. During the process the protein does not appear to bind to the resin. Give one possible explanation for why this is so.2. Explain how the decrease in salt concentration elites a protein from methylsepharose resin
3. What resin is a stronger binder of hydrophobic residues: methylsepharose or butylsepharose? Why? 1. You are trying to purify a protein that has a total ratio of hydrophobic to hydrophilic amino acids of 70:30 so you decide to purify the Lysate using HIC. During the process the protein does not appear to bind to the resin. Give one possible explanation for why this is so.
2. Explain how the decrease in salt concentration elites a protein from methylsepharose resin
3. What resin is a stronger binder of hydrophobic residues: methylsepharose or butylsepharose? Why?
2. Explain how the decrease in salt concentration elites a protein from methylsepharose resin
3. What resin is a stronger binder of hydrophobic residues: methylsepharose or butylsepharose? Why?
Explanation / Answer
1. HIC hydrophobic interaction Chromatography.
Since the protein has large proportion of hydrophobic groups it will not have affinity towards the resin. So it didn't bind to the resin.
2. Decreasing salt concentration will decrease hydrophobic interaction. Salt will increase the surface tension and hydrophobicity. That is high salt buffer will increase the solvation of protein. On decreasing salt concentration, the hydrophobic layer is exposed and forms an interaction with the resin.
3. Butyl sepharosr is stronger than methyl sepharose. Butyl resin is chemically more stable than methyl sepharose.
Related Questions
Navigate
Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.