4.Propose a two-step procedure to purify protein A from the othermolecules (there are multiple answers):Molecule"Protein A""Protein B""DNA C""DNA D"Molecular Weight(kDa)10.265.065.010.2pl8.08.0N/AN/Aa. Step 1: I would use(size, cation-exchange, or anion-exchange) chromatography using a pH (a number) buffer. In the spacebelow, sketch a chromatogram, label axes, and label what protein(s) arepresent in each peak. DO NOT WORRY ABOUT RELATIVE PEAK HEIGHTS.b. Step 2: I would take the(first, second, etc.) peak above and use(size, cation-exchange, or anion-exchange) chromatographyusing a pH (a number) buffer. In the space below, sketch achromatogram, label axes and label what protein(s) are present in eachpeak. DO NOT WORRY ABOUT RELATIVE PEAK HEIGHTS.

Ion exchange chromatography is a powerful separation technique used to purify proteins and other…

Answered: 4. Propose a two-step procedure to…

Biochemistry

9th Edition

ISBN:9781319114671

Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Chapter1: Biochemistry: An Evolving Science

Section: Chapter Questions

Problem 1P

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3. In what direction (toward the anode, toward the cathode, or toward neither) will the following proteins move in an electric field? a. Serum albumin (pl=4.9) at pH 8.0 b. Urease (pl=5.0) at pH3.0 and pH 9.0 c. Ribonuclease (pI=9.5) at pH 4.5, 9.5 and 11.0 d. Pepsin (pl=1.0) at pH 3.5, pH7.0 and pH 9.5
According to these data: Protein MW A. 125 43 55 C-7/D-48 C-7.0/D-7.5 92 3.9 C/D 11 F 14 6.5 C/D is a dimer What is the order of elution of a gel filtration column at pH= 8? Oal F. B. C/D, E, A OCFB. D. E. A OOAED, B. F. c OGAEC/D, B, F
14. A protein mixture consisting of proteins A, B, and C was subjected to various protein purification steps. First, the protein mixture was applied to an anion ion exchange column. When the column was washed with an increasing concentration gradient of chloride ions, the order of elution of the three proteins was B, then C, and finally A. Next, the protein mixture was chromatographed on a gel filtration column. The order of elution of the three proteins from this column was C, A, B. Finally, the protein mixture was passed over a chromatography column containing a Ni-NTA affinity matrix. Only protein A was retained by the column. Answer the following (i) Which protein (A, B, or C) has the highest and lowest positive charge (ii) Which protein has the highest and lowest mass (ii) Which protein has an affinity for the Ni-NTA matrix and why
5. Anesthetic gases used in surgery are known to bind to the hemoglobin molecule in red blood cells. The diagram below illustrates O2 binding curves of normal human HbA in the presence of the anesthetic gas dichloromethane (DCM). O UNTREATED 100 a Dсм 23 Torr Symbols: в Dсм 50 Torr x DCM I00 Torr o, 0 Torr DCM; O, 23 Torr DCM A, 50 Torr DCM x, 100 Torr DCM. 80 60 40 (1 Torr = 1 mm Hg.) 20 The solutions were buffered to pH 7.4. 0.5 1.0 1.5 2.0 log p02 (a) (* Hill plots. Label the axes and indicate on both the plot above and the Hill plot where the value of the dissociation equilibrium constant Ka for O2 binding is defined for 0 and 100 Torr of dichloromethane For the curves at 0 and 100 Torr of dichloromethane, draw their equivalent in the form of Kd Hb(O2)n Hb + nO2 % OXYGENATION
You are working as a Research Scientist I at Albany Molecular Research. For one of your bacterial growth media, you need to prepare 1.4 liters of a 35mM sucrose solution. What do you need to do? MW of sucrose is 342 g/mol Oa. dissolve 168 g of sucrose in ~ 1 liter and fill total volume to 1.4 liters O b. dissolve 8.55 g of sucrose in ~ 1 liter and fill total volume to 1.4 liters c. dissolve 16.8 g of sucrose in ~ 1 liter and fill total volume to 1.4 liters N d. dissolve 14.1 g of sucrose in~ 1 liter and fill total volume to 1.4 liters O e. dissolve 16.8 g of sucrose in 1.4 liters
14. Two ribosomal sub-units have sedimentation coefficients of S1 = 60 S and S2 = 40 S. Assuming that both sub-units have spherical shapes and their densities are equal, calculate an approximate ratio of their molecular weights (MW1/MW2) from their S values. SOLUTION:
which in is the best method for separating proteins A and B that have the properties below: Protein A: a tetramer of identical 80kDa subunits with a pl of 6.4 Protein B: a monomeric protein with a MW of 80kDa and a pl of 9.2 A.) Anion exchange chromatography at pH 11 B.) Gel exclusion chromatography C.) Cation exchange chromatography at pH 4 D. Cation exchange chromatography at pH 11 E.) SDS-PAGE
5. Anesthetic gases used in surgery are known to bind to the hemoglobin molecule in red blood cells. The diagram below illustrates O2 binding curves of normal human HbA in the presence of the anesthetic gas dichloromethane (DCM). O UNTREATED 100 Symbols: a Dсм 23 Torr • DCM 50 Torr x DCM I00 Torr o, 0 Torr DCM; О, 23 Torr DСМ Д, 50 Torr DСМ x, 100 Torr DCM. 80 60 40 (1 Torr = 1 mm Hg.) 20 The solutions were buffered to pH 7.4. 0.5 1.0 1.5 2.0 log p02 For the curves at 0 and 100 Torr of dichloromethane, draw their equivalent in the form of (a) (* Hill plots. Label the axes and indicate on both the plot above and the Hill plot where the value of the dissociation equilibrium constant Kd for O2 binding is defined for 0 and 100 Torr of dichloromethane Kd Hb(O2)n nO2 Hb + Be sure to label the axes, indicate the units on both axes, and make the Hill plot so that the values of Kd are equivalent to those on the saturation plot above. Indicate whether cooperativity, as indicated by the Hill…
Which of the following statements concerning SDS in SDS-PAGE is INCORRECT? a. SDS is a negatively charged detergent b. SDS can give all proteins a uniform charge c. the full name of SDS is sodium dodecyl sulfate d. SDS make protein gel electrophoresis to separate proteins based on molecular weight as well as protein surface charges e. SDS can denature proteins
1. In the SDS-PAGE figure below, looking at lane 6 there are two bands MamP, monomer and MamP dimer (disulfide bonded). Why does the MamP monomer run faster on the gel relative to the MamP dimer? 1 2 3 4 5 6 250 - 130 - 100 70 55 - Hisg MBPtev.Mamf MamP dimer 35 - 25- - MamP monomer 15 - 10 - If the gel was run under non-reducing conditions, would the pattern possibly be different and why? Molecular weight (kD)
23. In the structure below, the shaded areas H Ca Ca. H a) SDS gel electrophoresis b) mass spectrometry c) Edman degradation d) size exclusion chromatography Ca a) are unable to freely rotate around the C-N bond b) represent resonance structures c) represent a partial double bond between C and N d) all of the above Ca. 24. Which of the following protein separation techniques allows for the recovery of protein?

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