6DND

Crystal structure of wild-type (WT) human Glutamate oxaloacetate transaminase 1 (GOT1)

6DND の概要

• エントリーDOI: 10.2210/pdb6dnd/pdb
• 分子名称: Aspartate aminotransferase, cytoplasmic, TETRAETHYLENE GLYCOL, PYRIDOXAL-5'-PHOSPHATE, ... (4 entities in total)
• 機能のキーワード: aspartate aminotransferase, glutamate oxaloacetate transaminase 1, got1, plp, transferase
• 由来する生物種: Homo sapiens (Human)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 93076.81

構造登録者

Assar, Z.,Holt, M.C.,Stein, A.J.,Lairson, L.,Lyssiotis, C.A. (登録日: 2018-06-06, 公開日: 2018-11-14, 最終更新日: 2023-10-11)

主引用文献

Holt, M.C.,Assar, Z.,Beheshti Zavareh, R.,Lin, L.,Anglin, J.,Mashadova, O.,Haldar, D.,Mullarky, E.,Kremer, D.M.,Cantley, L.C.,Kimmelman, A.C.,Stein, A.J.,Lairson, L.L.,Lyssiotis, C.A.
Biochemical Characterization and Structure-Based Mutational Analysis Provide Insight into the Binding and Mechanism of Action of Novel Aspartate Aminotransferase Inhibitors.
Biochemistry, 57:6604-6614, 2018

PubMed Abstract: Pancreatic cancer cells are characterized by deregulated metabolic programs that facilitate growth and resistance to oxidative stress. Among these programs, pancreatic cancers preferentially utilize a metabolic pathway through the enzyme aspartate aminotransferase 1 [also known as glutamate oxaloacetate transaminase 1 (GOT1)] to support cellular redox homeostasis. As such, small molecule inhibitors that target GOT1 could serve as starting points for the development of new therapies for pancreatic cancer. We ran a high-throughput screen for inhibitors of GOT1 and identified a small molecule, iGOT1-01, with in vitro GOT1 inhibitor activity. Application in pancreatic cancer cells revealed metabolic and growth inhibitory activity reflecting a promiscuous inhibitory profile. We then performed an in silico docking analysis to study inhibitor-GOT1 interactions with iGOT1-01 analogues that possess improved solubility and potency properties. These results suggested that the GOT1 inhibitor competed for binding to the pyridoxal 5-phosphate (PLP) cofactor site of GOT1. To analyze how the GOT1 inhibitor bound to GOT1, a series of GOT1 mutant enzymes that abolished PLP binding were generated. Application of the mutants in X-ray crystallography and thermal shift assays again suggested but were unable to formally conclude that the GOT1 inhibitor bound to the PLP site. Mutational studies revealed the relationship between PLP binding and the thermal stability of GOT1 while highlighting the essential nature of several residues for GOT1 catalytic activity. Insight into the mode of action of GOT1 inhibitors may provide leads to the development of drugs that target redox balance in pancreatic cancer.

PubMed: 30365304
DOI: 10.1021/acs.biochem.8b00914

実験手法

X-RAY DIFFRACTION (2.1 Å)