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).
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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Please help. An explanation will be very helpful. Thank you
![Oxytocin is a 9-residue peptide hormone that is involved in signaling a number of pathways
related to reproduction and perinatal health. Like many peptide hormones, it is first produced as
a proprotein – a precursor of a protein that must be modified to produce the final, functional
protein. Oxytocin (OT) is initially produced as Oxytocin-neurophysin I (proOT-NPI). This is
processed by an enzyme into two proteins, prooxytocin (proOT) and Neurophysin I (NPI).
ProOT is then processed by another set of enzymes which removes a Lys-Arg dipeptide, yielding
active OT.
pro OT- NP I
Hz N f
Precessing Exzyme I
proOT
NP I
-cooo
Processing
I Enzymes 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 pI 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
OT
1007
N/A*
*OT is modified such that it neutral
except for one ionizable group, which is
the phenolic -OH of tyrosine (pKa=10).
1
+ : 0](/v2/_next/image?url=https%3A%2F%2Fcontent.bartleby.com%2Fqna-images%2Fquestion%2F64a87ed8-aba3-4af0-9288-25562d13f537%2F950498e0-5410-429d-aa31-8d4b896b4776%2Fq0c9w1_processed.jpeg&w=3840&q=75)
Transcribed Image Text:Oxytocin is a 9-residue peptide hormone that is involved in signaling a number of pathways
related to reproduction and perinatal health. Like many peptide hormones, it is first produced as
a proprotein – a precursor of a protein that must be modified to produce the final, functional
protein. Oxytocin (OT) is initially produced as Oxytocin-neurophysin I (proOT-NPI). This is
processed by an enzyme into two proteins, prooxytocin (proOT) and Neurophysin I (NPI).
ProOT is then processed by another set of enzymes which removes a Lys-Arg dipeptide, yielding
active OT.
pro OT- NP I
Hz N f
Precessing Exzyme I
proOT
NP I
-cooo
Processing
I Enzymes 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 pI 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
OT
1007
N/A*
*OT is modified such that it neutral
except for one ionizable group, which is
the phenolic -OH of tyrosine (pKa=10).
1
+ : 0
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