Date of Award

5-2016

Culminating Project Type

Thesis

Degree Name

Biological Sciences - Cell and Molecular: M.S.

Department

Biology

College

College of Science and Engineering

First Advisor

Bruce L. Jacobson

Second Advisor

Timothy Schuh

Third Advisor

Christopher Kvaal

Fourth Advisor

Latha Ramakrishnan

Creative Commons License

Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

Abstract

The fatty acid-binding proteins (FABPs) are a family of nine structurally related proteins, which bind to long-chain fatty acids with high affinity. Fatty acid binding proteins were first discovered in 1972, while conducting studies on the factors that regulate the uptake of intestinal fatty acids in rats. These proteins are most abundantly found in the tissues engaged in active lipid metabolism. Adipocyte fatty acid binding protein or aP2 is a member of this family, which is found in adipocytes and macrophages and integrates metabolic and inflammatory responses (Furuhashi et al., 2007).

Studies on aP2 knockout mice have shown that aP2 null mice were more insulin sensitive compared to their wild type counterparts when kept on a high fat diet. This lipid chaperone plays an important role in metabolic syndromes, including type 2 diabetes and atherosclerosis. Targeting aP2 with small-molecule inhibitors can lead to a new class of therapeutic agents that may provide a model of study to prevent metabolic diseases such as type 2 diabetes.

This study focuses on the purification of aP2 by ion exchange chromatography in preparation for co-crystallization with ibuprofen. Previous attempts to do this have failed due to the tight binding of fatty acid at the binding site of the protein. To overcome this issue, delipidation was attempted by means of chromatography using the Lipidex 1000 sorbent. The purified protein was crystallized using the hanging drop method with seeding in the presence of ibuprofen. This study describes the crystal structure of human aP2 with bound ibuprofen at 1.4 Å resolution. Crystalline 3

aP2 was orthorhombic with cell dimensions of a = 32.20 Å, b = 53.64 Å, c = 74.44 Å and the space group was P212121.

Comments/Acknowledgements

I would like to take this time to acknowledge and give thanks to several people who has helped me during my complete duration of Masters at St. Cloud State University. First of all, I would like to acknowledge my advisor Dr. Bruce Jacobson who trusted my decisions and provided me guidance, which is very much appreciated and welcomed. I would also like to thank my graduate committee: Dr. C. Kvall, Dr. Latha Ramakrishnan and Dr. T. Schuh. You all have supported me through those tough times when things were not good but I still was holding tight on my hopes. Thanks for cheering me and guiding up in those moments. I will like to thank my dear husband Shailesh Shrivastava. He supported me through this endeavor in every definition of the word. I thank you for that and everything else you have done to make it a success. I cannot forget to thank my dearest friend August, you not only walked with me to my final destination but also made the journey worthwhile, you are a true friend. I would like to thank my lab members who have helped me during the complete duration. I will like to thank my closest friend Smitha Balakrishnan who never lost faith in me I cherish your friendship and you will always be close to my heart. LastIy, I want to thank my parents Mr. Babu Lal Sharma and Ms. Tara Sharma for being there for me in good and bad times, your prayers have been answered and I will continue to grow and make you proud. My dearest brothers Arvind, Aditya and Arjun you are the anchors of my life thanks for always holding my hand I will never have enough words to thank you.

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