Thioesters are highly reactive centers for acyl-CoAs
which allows them to be utilized in a variety of differing enzyme chemistries.
As a result of this reactivity, structure-function studies of enzymes using acyl-CoA
substrates is difficult. When acyl-CoAs are used in structure-function studies,
they often result in a hydrolyzed CoA substrate fragment bound in the active
site or require only one of multiple substrates in order to be bound. This results
in a lack of information regarding enzyme interactions with the key thioester
and acyl chain. To overcome this challenging problem, I have synthesized
acetyl- and malonyl-CoA analogs where the thioester has been replaced by an ester
(oxygen), amide (nitrogen), or carbonyl (carbon) in a way that is easier,
cheaper, and more efficient than performed previously. In addition, we used our
synthetic analogs to study a enzymes which span different acyltransferase
mechanisms in a combination of kinetics and structure. With this work, it was
determined that the amide analogs were stable in all enzymes it was utilized
for, while the ester analogs were mostly stable, except the acetyl analog in
KasIII, where it acted as a pseudo substrate. As such, these synthetic analogs may
have future potential in either type of enzyme for structure-function studies,
albeit limited for the acetyl ester analog.
| Identifer | oai:union.ndltd.org:purdue.edu/oai:figshare.com:article/12724721 |
| Date | 29 July 2020 |
| Creators | Aaron B Benjamin (9175175) |
| Source Sets | Purdue University |
| Detected Language | English |
| Type | Text, Thesis |
| Rights | CC BY 4.0 |
| Relation | https://figshare.com/articles/thesis/IMPROVED_SYNTHESIS_OF_ACETYL-COA_AND_MALONYL-COA_ANALOGS_AND_THEIR_USE_TO_STUDY_STRUCTURE-FUNCTION_RELATIONSHIPS_OF_ACYLTRANSFERASES/12724721 |
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