Biochemistry: Concepts and Connections (2nd Edition)
2nd Edition
ISBN: 9780134641621
Author: Dean R. Appling, Spencer J. Anthony-Cahill, Christopher K. Mathews
Publisher: PEARSON
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Chapter 5, Problem 23P
If you want to purify a DNA-binding protein from a crude mixture of proteins at pH 7, should you use a DEAE-cellulose or a CM-cellulose column (see Figure 5.A5)? Briefly explain your reasoning.
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Chapter 5 Solutions
Biochemistry: Concepts and Connections (2nd Edition)
Ch. 5 - Prob. 1PCh. 5 - Draw the structure of the peptide DTLH, showing...Ch. 5 - Prob. 3PCh. 5 - Prob. 4PCh. 5 - Prob. 5PCh. 5 - Prob. 6PCh. 5 - Prob. 7PCh. 5 - Given the following peptide SEPIMAPVEYPK a....Ch. 5 - A mutant form of polypeptide hormone angiotensin...Ch. 5 - Prob. 10P
Ch. 5 - Prob. 11PCh. 5 - a. Write a possible sequence for an mRNA segment...Ch. 5 - 13. Assume the following portion of an mRNA Find a...Ch. 5 - Prob. 14PCh. 5 - Prob. 15PCh. 5 - Prob. 16PCh. 5 - Prob. 17PCh. 5 - Prob. 18PCh. 5 - You are a summer intern in a clinical hematology...Ch. 5 - Prob. 20PCh. 5 - Despite the fact that many peptides have critical...Ch. 5 - Based on the information in Figure 5.17, which...Ch. 5 - If you want to purify a DNA-binding protein from a...
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Need a deep-dive on the concept behind this application? Look no further. Learn more about this topic, biochemistry and related others by exploring similar questions and additional content below.Similar questions
- Suppose you have two genetic variants of a large protein that differ only inthat one contains a histidine (side chain pKa = 6.0) when the other has avaline (uncharged side chain).(a) Which would be better for separation: gel electrophoresis or isoelectricfocusing? Why?(b) What pH would you choose for the separation?arrow_forwardSpectroscopy is a useful tool to determine the concentration of DNA in a solution by measuring the UV absorbance at a wavelength of 260nm. When analyzing the purity of a DNA sample, an additional measurement of UV absorbance at 280nm is often taken to determine if proteins are is present as well. The ratio of 260/280 is then taken, and the closer this value is to 2 the more pure your DNA sample.a) Given that the aromatic rings present in the nitrogenous bases of DNA cause DNA to absorb UV light at 260nm, predict what might be responsible for proteins absorbing UV light at 280nm.b) You purify two DNA samples and measure the absorbance at 260nm and 280nm. For the first sample (Sample A) the absorbance at 280nm is 0, and for the second sample (Sample B) the absorbance at 260nm is 0.5. You are skeptical that Sample A is really that pure, and upon further testing you identify contaminating protein sequences (shown below) in both samples! Sample A contaminating protein: MSTSILEGAASTLSample B…arrow_forwardYou are performing an enzymatic reaction to digest DNA and nothing is working.The working solution for the digestion buffer should contain 400 mM Tris, pH 8.0 and 125 mM MgCl2. Mixed are: 0.2 ml of 2M Tris, pH 8.0 stock solution 0.25 ml of 5M MgCl2 stock solution *Solution completed with 0.55 ml water to make a final volume of 1ml. a) Before checking the calculation, what important question do you have to ask first for this specific solution? (not a generic question about all solutions) b) You are satisfied with the answer to a) and you believe he made a dilution error. You use the equation that applies to dilutions to check his calculations. Aha! you find the problem. What is it and how did you find out?arrow_forward
- You have begun your career in medicinal biochemistry and have just discovered a bacterial DNA plasmld (transferabl ring of DNA) that appears to destroy the Ebola virus. In order to characterize your new plasmid, the molar mass of the plasmid must be determined. You dissolve 25.00 mg of the purified plasmid in 0.200 mL of water at 2 °C and find the osmotlc pressure of this solution is 1.20 Torr at 20 °C and 1 atm pressure. Answer the following about the Ebola-killing plasmid. 33.) The osmotlc pressure of the system is: (a) 1 atm (b) 0.016 atm (c) 6.5 X 10-5 atm (d) 22.59 atm (e) 0.0016 atmarrow_forwardYou wish to purify a 55 kDa protein, but it is currently present as a mixture with another protein of approximately the same size. Briefly explain a purification strategy that you could use that would enable you to separate the two proteins.arrow_forwardThe following proteins were separated by SDS-PAGE in the presence of mercaptoethanol. Sketch the relative positions of the various polypeptides on the gel. Label the positive and negative ends of the gel.Protein A: 40 kDa single polypeptideProtein B: 80 kDa protein, made up of two subunits of molecular weight 20 kDa and 60 kDa, held together by noncovalent interactionsProtein C: 200 kDa protein, made up of four identical subunits (50 kDa each) linked together by disulfide bondsarrow_forward
- The OXA-M290 protein is next purified by size exclusion chromatography. To determine the best type of size exclusion resin to use, the size of OXA-M290 must first be determined. Earlier, you determined the amino acid sequence of OXA-M290 (MRVLALSAVFLVASIIGMPAVAKEWQENKSWNAHFTEHKSQGVVVLWNENKQQGFTNNLKRANQAFLPASSAKIPNSLIALDLGVVKDEHQVFKWDGQTRDIATWNRDHNLITAMKYSVVPVYQEFARQIGEARMSKMLHAFDYGNEDISGNVDSFWLDGGIRISATEQISFLRKLYHNKLHVSERSQRIVKQAMLTEANGDYIIRAKTGYDTKIGWWVGWVELDDNVWFFAMNMDMPTSDGLGLRQAITKEVLKQEKIIP). Based on the amino acid sequence, what is the molecular weight of this protein? You can use the free ProtParam tool (https://web.expasy.org/protparam/) to calculate the molecular weights of proteins. Make sure to include units in your answer. Note: The amino acid sequence reported earlier does not include the His-tag that was added to OXA-M290 by the pET-28a vector. However, you do not need to consider the amino acids in the His-tag in your answer to this question. For Context ONLY: For…arrow_forwardGiven the following eukaryotic DNA strand, transcribe and translate the DNA into a polypeptide using the 3’ – 5’ strand as the template. You may use drawings, diagrams, colours and annotations to describe how the DNA strand will be synthesized into a functional protein. (KEY: The letters SBMD are “made up” nucleic acids that depict non-coding regions in the DNA, hypothetically S pairs with B and M pairs with D).2.2. Describe what are missense mutations and its effects on structure and function using haemoglobin as an examplearrow_forward(a) Draw diagrams to show how the four synthetic oligonucleotides below could base-pair to form a stable model Holliday junction. W 5' GATCGCATTGTAGCCGTAGGTCCACTGTAA 3’ X 5' GTCCCATACGTAGCCGTAGGACATGTACCG 3' Y 5' CGGTACATGTCCTACGGCTACAATGCGATC 3' Z 5' TTACAGTGGACCTACGGCTACGTATGGGAC 3' I and 21arrow_forward
- Given the following eukaryotic DNA strand, transcribe and translate the DNA into a polypeptide using the 3’ – 5’ strand as the template. You may use drawings, diagrams, colours and annotations to describe how the DNA strand will be synthesized into a functional protein. (32) (KEY: The letters SBMD are “made up” nucleic acids that depict non-coding regions in the DNA, hypothetically S pairs with B and M pairs with D). 5’ - TATAAAAASSMSBMDATGSBDCCMBDBAATBSMDSTGTGTCCTMSBAG – 3’arrow_forwardDraw a structural formula for a nucleoside composed of the following. Q.) b-2-Deoxy-d-ribose and cytosinearrow_forwardAs described in the figure below, the technique of ultracentrifugation can be used to demonstrate whether a substance is composed of large covalently linked molecules, or smaller molecules held together in a complex by noncovalent bonds. You are studying two substances that both have very high molecular weights: hyaluronan, a carbohydrate found in the extracellular matrix with a MW of ~8 x 106 daltons, and the bacterial ribosome, with a MW of -2.5 x 106. Hyaluronan is a single, long-chain polymer, while the ribosome subunit is a complex of 55 different proteins plus three long RNA molecules. Which of the following describes the pattern you would expect to see from ultracentrifugation of these two substances? the sample loaded as a narrow band at the top of the tube (A) sample CENTRIFUGATION stabilizing sucrose gradient tube heterogeneous aggregates would sediment to produce a diffuse smear hemoglobin protein sediments as a single bandarrow_forward
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