(a)
Interpretation:
The digestion of octapeptide AVGWRVKS with trypsin. The most suitable method for separation of the products between ion-exchange or gel-filtration chromatography should be determined.
Concept introduction:
A peptide bond is present between the two amino acids in a protein. During the formation of a peptide bond, a molecule of water is released. The amino group of an amino acid gets associated with the carboxyl group of another. Polypeptides and proteins are the chains formed by the amino acids.
(b)
Interpretation:
If the protein was digested with chymotrypsin. The optimal separation method should be explained.
Concept introduction:
A peptide bond is present between the two amino acids in a protein. During the formation of a peptide bond, a molecule of water is released. The amino group of an amino acid gets associated with the carboxyl group of another. Polypeptides and proteins are the chains formed by the amino acids.
Want to see the full answer?
Check out a sample textbook solutionChapter 3 Solutions
BIOCHEM-ACHIEVE(FIRST DAY DISCOUNTED)
- Using a detailed scheme, propose a step-wise protocol to purify protein B by ion exchange chromatography (Explain your logic/choices).arrow_forward(b) Both laboratories used 10 micrograms of protein each in their kinetic assays. Protein concentrations weredetermined by the Bradford protein assay. Assay conditions employed in the two labs (pH, temperature,etc.) were also identical. What would be the most plausible cause for the discrepancy in the Vmax valuesfor the compound I? Explain.Recall that the Bradford assay measures total protein amounts in sample solution based on complexformation between a dye and proteins. Also, the assay solution used in both labs does not contain anyinhibitors.arrow_forwarda. Why proteins are extracted to a buffered solution? Briefly describe the components of ageneralized protein extraction buffer?.b. Describe the basis of affinity chromatography in protein purification.c. What is the most appropriate method of protein elution in affinity chromatography?d. List three examples of commonly employed combinations of ligand and protein in affinitychromatography.arrow_forward
- What are the advantages of gel filtration as a technique for protein purification? Identify two types of gels (sieving matrix) that may be used for gel filtration chromatography and discuss the types of samples which may be used for each. Discuss the principle behind affinity chromatography and differentiate it from gel filtration chromatography.arrow_forwarddetail how cation exchange chromatography works and what you would use to elute your target protein. What protein information would you need to facilitate this approach? Would you need to do any protein engineering to utilize cation exchange chromatography, justify your answer?arrow_forward(A) What property of a protein might make it difficult to transfer it from polyacrylamide gel to nitrocellulose? Explain your reasoning. (B) What parameter of the gel transfer protocol can be adjusted that might help improve the transfer of these problematic proteins to the nitrocellulose membrane? Explain your reasoning. (C) How can we check if proteins have been successfully transferred from the polyacrylamide gel to nitrocellulose?arrow_forward
- In a gel filtration chromatography, what type of gel must be used when the protein size is 2500 Da? Explain.arrow_forwardConsider the following properties of the protein components of a sample mixture as provided in the table below: 1. if the mixture is subjected to gel filtration chromotography which protein component elute first? 2. if the mixture is subjected to isoelectric focusing which protein will stop m oving nearest to the positive electrode? 3. if the mixture is subjected to cation-exchange chromotography using a buffer at ph 7 which protein will bind to the resin? 4.if the mixture is subjected to SDS-PAGE which protein will be at bottomost portion of gel? 5.if the mixture is subjected to hydrophobic interaction chromotography which protein will bind most strongly to the resin?arrow_forwardPlease provide a separation solution of combining two types of chromatography to isolate the protein A from the mixture containing protein A, B, C and D (shown as below). And then explain the related mechanisms in details. Protein A: Met-Leu-Leu-Leu-Leu-Val-Val-Val-lle-lle-lle-Leu-Leu-Leu-Leu- Protein B: Met-Asp-Glu-Glu-Glu-Glu-Asp-Asp-Asp-Glu-Glu-Asp-Asp-Glu Protein C: Met-Leu-Leu-Leu-Leu-Val-Val-Val-lle-lle-lle-Leu-Leu-Leu-Leu-Val-Val-Val- Ile-lle-lle-Leu-Leu- Leu-Leu-Val-Val-Val-lle-lle-lle-Leu-Leu-Leu-Leu-Val-Val-Val-lle-lle- lle-Leu-Leu-Leu-Leu-Val-Val-Val-Ile- Protein D: Met-Asp-Glu--Glu-Glu-Glu-Asp-Asp-Asp-Glu-Glu-Asp-Asp-Glu-Glu-Asp- Glu-Asp-Glu-Glu-Asp-Asp-Glu-Glu-Glu-Glu-Asp-Asp-Asp-Glu-Glu-Asp-Asp-Glu-Glu- Asp-Glu-Asp-Glu-Glu-Asp-Glu-Glu-Asparrow_forward
- Which technique will be most suitable for desalting the protein or analyzing the actual dimensional (D) native form and what would be the advantages of this selection? Discuss two to three supportive evidence of the technique application.arrow_forwardUnder most in vitro assay conditions, the enzyme is used in catalytic amounts (10 to 10" M). Estimate the concentration of an enzyme in a living cell. Assume that (a) fresh tissue is 80% water and all of it is intracellular, (b) the total soluble protein in a cell represents 15% of the wet weight, (c) all the soluble proteins are enzymes, (d) the average molecular weight of a protein is 150,000, and (e) about 1000 different enzymes are present.arrow_forwardGel-filtration chromatography is a useful method for removing salts, such as ammonium sulfate, from protein solutions. Describe how such a separation is accomplished.arrow_forward
- BiochemistryBiochemistryISBN:9781319114671Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.Publisher:W. H. FreemanLehninger Principles of BiochemistryBiochemistryISBN:9781464126116Author:David L. Nelson, Michael M. CoxPublisher:W. H. FreemanFundamentals of Biochemistry: Life at the Molecul...BiochemistryISBN:9781118918401Author:Donald Voet, Judith G. Voet, Charlotte W. PrattPublisher:WILEY
- BiochemistryBiochemistryISBN:9781305961135Author:Mary K. Campbell, Shawn O. Farrell, Owen M. McDougalPublisher:Cengage LearningBiochemistryBiochemistryISBN:9781305577206Author:Reginald H. Garrett, Charles M. GrishamPublisher:Cengage LearningFundamentals of General, Organic, and Biological ...BiochemistryISBN:9780134015187Author:John E. McMurry, David S. Ballantine, Carl A. Hoeger, Virginia E. PetersonPublisher:PEARSON