Keywords
acetyl CoA carboxylase, malonyl CoA, isostere
Select the category the research project fits.
Life Sciences
Is this submission part of ICaP/PW (Introductory Composition at Purdue/Professional Writing)?
No
Abstract
Malonyl-CoA is a molecule involved in a variety of important reactions and it is biologically important as it is required for fatty acid synthesis. This compound is also highly reactive making it and the enzymes that use it difficult to study. As a result, in order to learn mechanistic information, more stable analogs of malonyl-CoA were made in order to mimic the size, shape and acidity of the thioester and carboxylate groups. Throughout the semester, some malonyl-CoA analogs were synthesized and bacterial inhibition growth assays were performed with Escherichia Coli, Bacillus cereus, and Micrococcus luteus. However, the inhibition predicted was not seen from any of the analogs previously synthesized. As a result, it was concluded that the information suggesting that the analogs did inhibit bacterial growth was not replicable and no minimal inhibition concentration against these microorganisms was able to be determined. To continue this research, more work will be done on synthesis of analogs that replace thioester sulfur with a carbon atom. Currently, we have synthesized several intermediate steps to completing analogs containing a ketone with different carboxylate bioisosteres. These analogs will be used to study different acyl-CoA utilizing enzymes by co-crystallizing the analogs with enzymes such as acetyl-CoA carboxylase (ACC). Finally, the Kd of the different analogs with ACC will be determined to observe how tightly they bind in the enzyme active site. We will measure this value using either isothermal titration calorimetry (ITC) or microscale thermophoresis (MST). This kinetic data in combination with the structural data gained through X-ray crystallography will allow us to determine catalytic details of different enzymes, such as ACC.
Recommended Citation
Pascual-Garrigos, Ana, "Understanding the Transcarboxylation Reaction Performed by Acetyl CoA Carboxylase" (2019). Purdue Undergraduate Research Conference. 2.
https://docs.lib.purdue.edu/purc/2019/Posters/2
Understanding the Transcarboxylation Reaction Performed by Acetyl CoA Carboxylase
Malonyl-CoA is a molecule involved in a variety of important reactions and it is biologically important as it is required for fatty acid synthesis. This compound is also highly reactive making it and the enzymes that use it difficult to study. As a result, in order to learn mechanistic information, more stable analogs of malonyl-CoA were made in order to mimic the size, shape and acidity of the thioester and carboxylate groups. Throughout the semester, some malonyl-CoA analogs were synthesized and bacterial inhibition growth assays were performed with Escherichia Coli, Bacillus cereus, and Micrococcus luteus. However, the inhibition predicted was not seen from any of the analogs previously synthesized. As a result, it was concluded that the information suggesting that the analogs did inhibit bacterial growth was not replicable and no minimal inhibition concentration against these microorganisms was able to be determined. To continue this research, more work will be done on synthesis of analogs that replace thioester sulfur with a carbon atom. Currently, we have synthesized several intermediate steps to completing analogs containing a ketone with different carboxylate bioisosteres. These analogs will be used to study different acyl-CoA utilizing enzymes by co-crystallizing the analogs with enzymes such as acetyl-CoA carboxylase (ACC). Finally, the Kd of the different analogs with ACC will be determined to observe how tightly they bind in the enzyme active site. We will measure this value using either isothermal titration calorimetry (ITC) or microscale thermophoresis (MST). This kinetic data in combination with the structural data gained through X-ray crystallography will allow us to determine catalytic details of different enzymes, such as ACC.