(b) To the right is a diagram of the chemical groups in carboxymethyl-cellulose and DEAE-cellulose that are used for ion exchange chromatography to purify proteins. At pH 8 which chromatographic resin has to be used to separate iso-1-cytochrome c from iso-2- cytochrome c from a lysate of yeast cells? If the proteins are separated by application of a concentration gradient carboxymethyl of NaCl applied to the column, which ion Na® or Cle dis- places the proteins from the binding sites on the chroma- tographic resin? cellulose OH H3C CH3 ΘΝΗ HO HỌC, NH H3C 2-(diethylamino)ethyl cellulose (DEAE cellulose) Which protein iso-1-cytochrome c or iso-2-cytochrome c will be the first to be eluted from the column and why? Both cytochromes in their ferrous form are substrates of yeast cytochrome c peroxidase (CCP) and react optimally with this enzyme at pH 8 forming a Michaelis complex of 1:1 stoichiometry (CcP:cyt c). If the isoionic point of the peroxidase is 5.25, what must be the overall electrostatic charge on the peroxidase enzyme for this reaction to occur, positive or negative?
(b) To the right is a diagram of the chemical groups in carboxymethyl-cellulose and DEAE-cellulose that are used for ion exchange chromatography to purify proteins. At pH 8 which chromatographic resin has to be used to separate iso-1-cytochrome c from iso-2- cytochrome c from a lysate of yeast cells? If the proteins are separated by application of a concentration gradient carboxymethyl of NaCl applied to the column, which ion Na® or Cle dis- places the proteins from the binding sites on the chroma- tographic resin? cellulose OH H3C CH3 ΘΝΗ HO HỌC, NH H3C 2-(diethylamino)ethyl cellulose (DEAE cellulose) Which protein iso-1-cytochrome c or iso-2-cytochrome c will be the first to be eluted from the column and why? Both cytochromes in their ferrous form are substrates of yeast cytochrome c peroxidase (CCP) and react optimally with this enzyme at pH 8 forming a Michaelis complex of 1:1 stoichiometry (CcP:cyt c). If the isoionic point of the peroxidase is 5.25, what must be the overall electrostatic charge on the peroxidase enzyme for this reaction to occur, positive or negative?
Biochemistry
6th Edition
ISBN:9781305577206
Author:Reginald H. Garrett, Charles M. Grisham
Publisher:Reginald H. Garrett, Charles M. Grisham
Chapter18: Glycolysis
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Problem 14P: Energetic of Fructose-1 ,6-bis P Hydrolysis (Integrates with Chapter 3.) The standard free energy...
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![2. In Bakers yeast there are two variants of cytochrome c, a heme containing protein of 108 amino
acid residues that catalyzes electron transfer reactions. The two forms of cytochrome c known as
iso-1 and iso-2 cytochrome c differ in 17 of the 108 amino acids. The content of proton dis-sociable
side chains for the two cytochromes c is given in the table below:
Side chain
pKa iso-1 cytochrome c Ziso-1
Arginine
Aspartate
Cysteine
Glutamate
Histidine
Lysine
Tyrosine
-NH3Ⓡ
-COOH
12.5
4.0
8.0
4.0
6.0
10.0
9.0
7.0
4.0
34
1
4
4
16
⑤
5
1
1
iso-2 cytochrome c
(b) To the right is a diagram of the chemical
groups in carboxymethyl-cellulose and DEAE-cellulose
that are used for ion exchange chromatography to purify
proteins. At pH 8 which chromatographic resin has to be
used to separate iso-1-cytochrome c from iso-2-
cytochrome c from a lysate of yeast cells? If the proteins
are separated by application of a concentration gradient carboxymethyl
of NaCl applied to the column, which ion Na® or Cle dis-
places the proteins from the binding sites on the chroma-
tographic resin?
Joo
cellulose
OH
3
77751 vouw
5
7
3
H3C
CH 3
NH
HO
Ziso-2
HỌC LẠNH
H3C
2-(diethylamino)ethyl
cellulose
(DEAE cellulose)
Which protein iso-1-cytochrome c or iso-2-cytochrome c will be the first to be eluted from the
column and why?
Both cytochromes in their ferrous form are substrates of yeast cytochrome c peroxidase (CCP) and
react optimally with this enzyme at pH 8 forming a Michaelis complex of 1:1 stoichiometry (CcP:cyt
c). If the isoionic point of the peroxidase is 5.25, what must be the overall electrostatic charge
on the peroxidase enzyme for this reaction to occur, positive or negative?](/v2/_next/image?url=https%3A%2F%2Fcontent.bartleby.com%2Fqna-images%2Fquestion%2Fc86ae8b4-8072-44c1-858a-e689a302355a%2F03f686df-693f-4f28-a1fa-362c9ff03df1%2Fdq2amzs_processed.png&w=3840&q=75)
Transcribed Image Text:2. In Bakers yeast there are two variants of cytochrome c, a heme containing protein of 108 amino
acid residues that catalyzes electron transfer reactions. The two forms of cytochrome c known as
iso-1 and iso-2 cytochrome c differ in 17 of the 108 amino acids. The content of proton dis-sociable
side chains for the two cytochromes c is given in the table below:
Side chain
pKa iso-1 cytochrome c Ziso-1
Arginine
Aspartate
Cysteine
Glutamate
Histidine
Lysine
Tyrosine
-NH3Ⓡ
-COOH
12.5
4.0
8.0
4.0
6.0
10.0
9.0
7.0
4.0
34
1
4
4
16
⑤
5
1
1
iso-2 cytochrome c
(b) To the right is a diagram of the chemical
groups in carboxymethyl-cellulose and DEAE-cellulose
that are used for ion exchange chromatography to purify
proteins. At pH 8 which chromatographic resin has to be
used to separate iso-1-cytochrome c from iso-2-
cytochrome c from a lysate of yeast cells? If the proteins
are separated by application of a concentration gradient carboxymethyl
of NaCl applied to the column, which ion Na® or Cle dis-
places the proteins from the binding sites on the chroma-
tographic resin?
Joo
cellulose
OH
3
77751 vouw
5
7
3
H3C
CH 3
NH
HO
Ziso-2
HỌC LẠNH
H3C
2-(diethylamino)ethyl
cellulose
(DEAE cellulose)
Which protein iso-1-cytochrome c or iso-2-cytochrome c will be the first to be eluted from the
column and why?
Both cytochromes in their ferrous form are substrates of yeast cytochrome c peroxidase (CCP) and
react optimally with this enzyme at pH 8 forming a Michaelis complex of 1:1 stoichiometry (CcP:cyt
c). If the isoionic point of the peroxidase is 5.25, what must be the overall electrostatic charge
on the peroxidase enzyme for this reaction to occur, positive or negative?
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