proOT NP I J Processing V Eneymes Ž:3 OT You have been tasked to purify mature OT from a cell lysate expressing proOT-NPI that has been tagged with a polyhistidine tag at the C-terminus. Design a purification scheme to accomplish this goal. You have all of the biochemical equipment we've discussed in class, as well as purified Processing Enzymes 1, 2, and 3. The MW and pl values for proOT-NPI, NPI, proOT, and OT are provided in the table to the right. You should assume conversion reactions will not go to completion, so you will need to purify any products from the starting material. Assume that compounds must be 1000 Da different to separate with size- exclusion chromatrography and have a pl difference of 1 to separate via ion exchange chromatography. Your answer can include a drawing, but include some text so that I can understand your rationale for each step. Protein MW (Da) pl ProOT-NPI 10593 4.94 NPI 9600 4.94 ProOT 1010 5.51 от 1007 N/A* *OT is modified such that it neutral except for one ionizable group, which is the phenolic -OH of tyrosine (pK=10).

proOT NP I J Processing V Eneymes Ž:3 OT You have been tasked to purify mature OT from a cell lysate expressing proOT-NPI that has been tagged with a polyhistidine tag at the C-terminus. Design a purification scheme to accomplish this goal. You have all of the biochemical equipment we've discussed in class, as well as purified Processing Enzymes 1, 2, and 3. The MW and pl values for proOT-NPI, NPI, proOT, and OT are provided in the table to the right. You should assume conversion reactions will not go to completion, so you will need to purify any products from the starting material. Assume that compounds must be 1000 Da different to separate with size- exclusion chromatrography and have a pl difference of 1 to separate via ion exchange chromatography. Your answer can include a drawing, but include some text so that I can understand your rationale for each step. Protein MW (Da) pl ProOT-NPI 10593 4.94 NPI 9600 4.94 ProOT 1010 5.51 от 1007 N/A* *OT is modified such that it neutral except for one ionizable group, which is the phenolic -OH of tyrosine (pK=10).

Human Heredity: Principles and Issues (MindTap Course List)

11th Edition

ISBN:9781305251052

Author:Michael Cummings

Publisher:Michael Cummings

Chapter10: From Proteins To Phenotypes

Section: Chapter Questions

Problem 13QP: Suppose that in the formation of phenylalanine hydroxylase mRNA, the exons of the pre-mRNA fail to...

See similar textbooks

Similar questions

A mutant form of polypeptide hormone angiotensin II has the amino acidcomposition (Asp, Arg, Ile, Met, Phe, Pro, Tyr, Val)The following observations are made:• Trypsin yields a dipeptide containing Asp and Arg and a hexapeptide withall the rest.• Cyanogen bromide cleavage yields a dipeptide containing Phe and Proand a hexapeptide containing all the others.• Chymotrypsin cleaves the hormone into two tetrapeptides, of composition (Asp, Arg, Tyr, Val) and (Ile, Met, Phe, Pro)• The dipeptide of composition (Pro, Phe) cannot be cleaved by either chymotrypsin or carboxypeptidase.What is the sequence of angiotensin II?
A study is conducted to assess the molecular mechanisms of hormone X. A radioactively labeled form of this hormone is incubated with human cells in tissue culture for 30 minutes, and then incubated briefly with a cross-linking agent. The plasma membrane, cytosolic, and nuclear fractions are isolated from the cell lysate, and the subcellular proteins are separated from these fractions by gel electrophoresis. The only protein bound to the labeled hormone is identified in the new clear fraction. Hormone X is most likely which of the following? Epinephrine Glucagon Growth hormone Insulin Thyroxine
In response to a hormone secreted by a cell of the opposite mating type, a yeast cell undergoes a complex series of physiological changes involving the activity of about 200 genes and cytoplasmic proteins. They include blockingDNA synthesis, growing toward the mating partner, fusion of the plasma membranes of the two cells, and fusion of their nuclear membranes. Explain how all these events can be controlled through a complex signaling cascade that is triggered by the binding of the hormone to a G protein–linkedreceptor.
TPCK and TLCK are irreversible inhibitors of serine proteases. One ofthese inhibits trypsin and the other chymotrypsin. Which is which? Explainyour reasoning. Suggest the effects of each of the following mutations on the physiologicalrole of chymotrypsinogen:(a) R15S(b) C1S(c) T147S
Describe the intracellular, biochemical downstream signal cascade that results from the binding of menthol to TRPM8
HbA1c is used to monitor blood glucose levels because hemoglobin is the only protein in blood that is covalently modified by glucose. True False Insulin Glargine is a long-acting form of insulin that is synthesized with several D-amino acids that slow its proteolytic degradation and extend the half-life of the insulin Glargine molecule. True False
A mutant form of polypeptide hormone has the following amino acid composition: Asp, Arg, lle, Met, Phe, Pro, Tyr, Val The following observations are made: Trypsin yields a dipeptide containing Asp and Arg, and a hexapeptide with all the rest. Cyanogen bromide cleavage yields a dipeptide containing Phe and Pro, and a hexapeptide containing all the others. Chymotrypsin cleaves the hormone into two tetrapeptides with the following composition: (Asp, Arg, Tyr, Val) and (lle, Met Phe, Pro) What is the amino acid sequence of this polypeptide?
Ligand binding to proteins may occur with varying strengths; some ligands bind tightly to proteins while others bind less tightly. The strengths of reversible binding are determined experimentally by varying concentrations of ligands, and measuring the saturation of the protein in the various ligand concentrations. One such laboratory study investigated the binding of a hormone to three different receptor proteins in the cell membrane. The data collected are shown in the table below 1) Which of the proteins demonstrate the tightest/strongest binding of the hormone? Which demonstrates the least tight binding of the hormone? What is the basis for your answer?
explain the following prperties of G protein: structure of G- activation cycle and signaling pathway for Gaq
The protein fragments ABS1 and ABS2 of tropomodulin were produced as fusion proteins with chitin binding domain and purified by a chitin column. Explain the principles of this type of affinity chromatography and support it with a self‐drawn figure. This is with regards to Tropomodulin/ F-Actin complex.
A mutant form of polypeptide hormone has the following amino acid composition: Asp, Arg, lle, Met, Phe, Pro, Tyr. Val The following observations are made: Trypsin yields a dipeptide containing Asp and Arg, and a hexapeptide with all the rest. Cyanogen bromide cleavage yields a dipeptide containing Phe and Pro, and a hexapeptide containing all the others. Chymotrypsin cleaves the hormone into two tetrapeptides with the following composition: (Asp, Arg, Tyr, Val) and (lle, Met Phe, Pro) The dipeptide of composition (Pro, Phe) cannot be cleaved by either chymotrypsin or carboxypeptidase. What is the amino acid sequence of this polypeptide?
One such laboratory study investigated the binding of a hormone to three different receptor proteins in the cell membrane. The data collected are shown in the table below 1) Provide a brief explanation as to why ligand binding toproteins must be a reversible process. 2) Calculate the dissociation constant (Kd) for the hormone binding to each of the three proteins.