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A human fatty acid synthase inhibitor binds β-ketoacyl reductase in the keto-substrate site


Human fatty acid synthase (hFAS) is a complex, multifunctional enzyme that is solely responsible for the de novo synthesis of long chain fatty acids. hFAS is highly expressed in a number of cancers, with low expression observed in most normal tissues. Although normal tissues tend to obtain fatty acids from the diet, tumor tissues rely on de novo fatty acid synthesis, making hFAS an attractive metabolic target for the treatment of cancer. We describe here the identification of GSK2194069, a potent and specific inhibitor of the β-ketoacyl reductase (KR) activity of hFAS; the characterization of its enzymatic and cellular mechanism of action; and its inhibition of human tumor cell growth. We also present the design of a new protein construct suitable for crystallography, which resulted in what is to our knowledge the first co-crystal structure of the human KR domain and includes a bound inhibitor.

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Figure 1: Mechanism of inhibition of GSK2194069 versus hFAS KR.
Figure 2: GSK2194069 inhibits FAS in cancer cell lines.
Figure 3: Design of the hFAS tridomain construct and comparison with the pig FAS crystal structure.
Figure 4: GSK2194069 binds in the active site and at the domain interface.

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Primary accessions

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Referenced accessions

NCBI Reference Sequence

Protein Data Bank


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We are grateful to C. Aquino for designing and first preparing GSK2194069 and to A. Chaudhari, J. Mack and K. Erhard for preparing additional quantities. We thank Y. Shen for construct engineering, J. Christie for virus generation and G. Barrett for large-scale cell culture. We thank B. Rodríguez, M. Jesús Vázquez and R. Liu for the high-throughput screening data; K. McKenzie and J. Gross for the initial FAS inhibition data for GSK2194069; and G. Waitt for the LC/MS/MS analyses in the M-CoA experiments. We also thank G. Graczyk-Millbrandt and M. Wang for final structure confirmation by NMR. Support of this program by R. Wooster is gratefully acknowledged. Use of the Advanced Photon Source was supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences, under contract no. DE-AC02-06CH11357. Use of the Life Sciences Collaborative Access Team (LS-CAT) Sector 21 was supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (grant no. 085P1000817).

Author information




M.L.M., S.P.W. and K.K.B. designed constructs. L.W. performed and interpreted crystallization studies. S.P.W. solved and analyzed structures. W.A.B. coordinated cell preparation and purified all of the proteins. A.R.R. and R.D.T. performed biochemical mechanism studies. J.A.K. performed and analyzed western blotting and performed M-CoA experiments. R.N.P. performed and analyzed cellular growth assays and prepared cells for NMR analysis. J.B. performed and analyzed NMR studies. C.A.P. selected 2. A.R.R., G.Z., R.D.T. and M.A.H. designed experiments. A.R.R., G.Z., R.D.T., M.L.M., C.A.P. and M.A.H. interpreted and/or analyzed data. A.R.R., S.P.W., M.L.M., M.A.H. and C.A.P. wrote the paper, which was reviewed by all of the authors.

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Correspondence to Cynthia A Parrish.

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All of the authors are employees of GlaxoSmithKline, and all are or have been shareholders of GlaxoSmithKline.

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Supplementary Results, Supplementary Tables 1–3 and Supplementary Figures 1–7. (PDF 837 kb)

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Hardwicke, M., Rendina, A., Williams, S. et al. A human fatty acid synthase inhibitor binds β-ketoacyl reductase in the keto-substrate site. Nat Chem Biol 10, 774–779 (2014).

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