In regards to question number two, I could not find anywhere in the paper where they discusses this aspect; however, I think that this column could be used for cytochrome c analysis of multiple species. The similarity between human cytochrome c and other species is probably pretty high and I would imagine that the aptamer binds to a common site found in the protein across organisms. For instance, I used expasy to run a sequence similarity of cytochrome c for homo sapiens and drosophila melanogaster (fruit fly) and the result is that they are about 79% similar.
I think the apatamer used in this monolithic column probably binds to a common protein region and thus can be used for multiple organisms.
I am not entirely sure what sort of information could be gleaned by using this monolithic column for use in other species. While I am sure that this column could be used for separation and isolation of cytochrome c through multiple species, the binding constants determined would have very little meaning.
For question 1, the derivation was supplied in the supplementary information.
I think if different species of cytochrome c can be retained on the column, the binding constants would have some meaning. If two species genomes have a high degree of similarity yet their binding contants are the same, then there is not a whole lot of change obviously. If their is a different binding constant with similar genomes, this could correlate to post-translational modifications or changes in protein folding.
Question 2
I agree with Josh, if the cytochrome c obtained from a different species is similar enough to the cytochrome c used in the study then the column should have no problem with the separation. It possible that the other cytochome c might bind with a different affinity and separations of cytochrome c from two species might be possible as well
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Comments
1. The main manuscript does not show the derivation of the equation used to calculate the Kd and the saturation constant. Please derive.
2. Can the monolithic column be used for the separation and isolation of cytochrome c obtained from different species?
Posted by: Edgar Arriaga | April 23, 2008 12:42 AM
In regards to question number two, I could not find anywhere in the paper where they discusses this aspect; however, I think that this column could be used for cytochrome c analysis of multiple species. The similarity between human cytochrome c and other species is probably pretty high and I would imagine that the aptamer binds to a common site found in the protein across organisms. For instance, I used expasy to run a sequence similarity of cytochrome c for homo sapiens and drosophila melanogaster (fruit fly) and the result is that they are about 79% similar.
I think the apatamer used in this monolithic column probably binds to a common protein region and thus can be used for multiple organisms.
Posted by: Josh Ochocki | April 23, 2008 10:47 AM
I am not entirely sure what sort of information could be gleaned by using this monolithic column for use in other species. While I am sure that this column could be used for separation and isolation of cytochrome c through multiple species, the binding constants determined would have very little meaning.
Posted by: Eric Olson | April 25, 2008 05:11 PM
For question 1, the derivation was supplied in the supplementary information.
I think if different species of cytochrome c can be retained on the column, the binding constants would have some meaning. If two species genomes have a high degree of similarity yet their binding contants are the same, then there is not a whole lot of change obviously. If their is a different binding constant with similar genomes, this could correlate to post-translational modifications or changes in protein folding.
Posted by: Chad Satori | April 27, 2008 10:54 PM
Answer to Question 1
Download file
Question 2
I agree with Josh, if the cytochrome c obtained from a different species is similar enough to the cytochrome c used in the study then the column should have no problem with the separation. It possible that the other cytochome c might bind with a different affinity and separations of cytochrome c from two species might be possible as well
Posted by: Craig Bishop | April 28, 2008 09:57 AM