Abstract
De novo synthesis of fatty acids in the cytosol of animal cells is carried out by the multifunctional, homodimeric fatty acid synthase (FAS). Cryo-EM analysis of single FAS particles imaged under conditions that limit conformational variability, combined with gold labeling of the N termini and structural analysis of the FAS monomers, reveals two coiled monomers in an overlapping arrangement. Comparison of dimeric FAS structures related to different steps in the fatty acid synthesis process indicates that only limited local rearrangements are required for catalytic interaction among different functional domains. Monomer coiling probably contributes to FAS efficiency and provides a structural explanation for the reported activity of a FAS monomer dimerized to a catalytically inactive partner. The new FAS structure provides a new paradigm for understanding the architecture of FAS and the related modular polyketide synthases.
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References
Alberts, A.W., Strauss, A.W., Hennessy, S. & Vagelos, P.R. Regulation of synthesis of hepatic fatty acid synthetase: binding of fatty acid synthetase antibodies to polysomes. Proc. Natl. Acad. Sci. USA 72, 3956–3960 (1975).
Stoops, J.K. et al. Presence of two polypeptide chains comprising fatty acid synthetase. Proc. Natl. Acad. Sci. USA 72, 1940–1944 (1975).
Smith, S., Agradi, E., Libertini, L. & Dileepan, K.N. Specific release of the thioesterase component of the fatty acid synthetase multienzyme complex by limited trypsinization. Proc. Natl. Acad. Sci. USA 73, 1184–1188 (1976).
Wakil, S.J. Fatty acid synthase, a proficient multifunctional enzyme. Biochemistry 28, 4523–4530 (1989).
Smith, S., Witkowski, A. & Joshi, A.K. Structural and functional organization of the animal fatty acid synthase. Prog. Lipid Res. 42, 289–317 (2003).
Chirala, S.S. et al. Fatty acid synthesis is essential in embryonic development: fatty acid synthase null mutants and most of the heterozygotes die in utero. Proc. Natl. Acad. Sci. USA 100, 6358–6363 (2003).
Rufo, C. et al. Involvement of a unique carbohydrate-responsive factor in the glucose regulation of rat liver fatty-acid synthase gene transcription. J. Biol. Chem. 276, 21969–21975 (2001).
Loftus, T.M. et al. Reduced food intake and body weight in mice treated with fatty acid synthase inhibitors. Science 288, 2379–2381 (2000).
Furuya, Y., Akimoto, S., Yasuda, K. & Ito, H. Apoptosis of androgen-independent prostate cell line induced by inhibition of fatty acid synthesis. Anticancer Res. 17, 4589–4593 (1997).
Kuhajda, F.P. et al. Synthesis and antitumor activity of an inhibitor of fatty acid synthase. Proc. Natl. Acad. Sci. USA 97, 3450–3454 (2000).
De Schrijver, E., Brusselmans, K., Heyns, W., Verhoeven, G. & Swinnen, J.V. RNA interference-mediated silencing of the fatty acid synthase gene attenuates growth and induces morphological changes and apoptosis of LNCaP prostate cancer cells. Cancer Res. 63, 3799–3804 (2003).
Chirala, S.S., Jayakumar, A., Gu, Z.W. & Wakil, S.J. Human fatty acid synthase: role of interdomain in the formation of catalytically active synthase dimer. Proc. Natl. Acad. Sci. USA 98, 3104–3108 (2001).
Smith, S. The animal fatty acid synthase: one gene, one polypeptide, seven enzymes. FASEB J. 8, 1248–1259 (1994).
Stoops, J.K. & Wakil, S.J. Animal fatty acid synthetase. A novel arrangement of the β-ketoacyl synthetase sites comprising domains of the two subunits. J. Biol. Chem. 256, 5128–5133 (1981).
Stoops, J.K. & Wakil, S.J. Animal fatty acid synthetase. Identification of the residues comprising the novel arrangement of the β-ketoacyl synthetase site and their role in its cold inactivation. J. Biol. Chem. 257, 3230–3235 (1982).
Joshi, A.K., Rangan, V.S. & Smith, S. Differential affinity labeling of the two subunits of the homodimeric animal fatty acid synthase allows isolation of heterodimers consisting of subunits that have been independently modified. J. Biol. Chem. 273, 4937–4943 (1998).
Rangan, V.S., Joshi, A.K. & Smith, S. Mapping the functional topology of the animal fatty acid synthase by mutant complementation in vitro. Biochemistry 40, 10792–10799 (2001).
Witkowski, A. et al. Dibromopropanone cross-linking of the phosphopantetheine and active-site cysteine thiols of the animal fatty acid synthase can occur both inter- and intrasubunit. Reevaluation of the side-by-side, antiparallel subunit model. J. Biol. Chem. 274, 11557–11563 (1999).
Joshi, A.K., Rangan, V.S., Witkowski, A. & Smith, S. Engineering of an active animal fatty acid synthase dimer with only one competent subunit. Chem. Biol. 10, 169–173 (2003).
Kitamoto, T., Nishigai, M., Sasaki, T. & Ikai, A. Structure of fatty acid synthetase from the Harderian gland of guinea pig. Proteolytic dissection and electron microscopic studies. J. Mol. Biol. 203, 183–195 (1988).
Brink, J. et al. Quaternary structure of human fatty acid synthase by electron cryomicroscopy. Proc. Natl. Acad. Sci. USA 99, 138–143 (2002).
Wakil, S.J., Stoops, J.K. & Joshi, V.C. Fatty acid synthesis and its regulation. Annu. Rev. Biochem. 52, 537–579 (1983).
Frank, J. Three-Dimensional Electron Microscopy of Macromolecular Assemblies 342 (Academic Press, San Diego, 1996).
Brink, J. et al. Experimental verification of conformational variation of human fatty acid synthase as predicted by normal mode analysis. Structure 12, 185–191 (2004).
Radermacher, M. The three-dimensional reconstruction of single particles from random and non-random tilt series. J. Electron Microsc. Tech. 9, 359–394 (1988).
Dubochet, J. et al. Cryo-electron microscopy of vitrified specimens. Q. Rev. Biophys. 21, 129–228 (1988).
Penczek, P., Grassucci, R.A. & Frank, J. The ribosome at improved resolution: new techniques for merging and orientation refinement in 3D cryo-electron microscopy of biological particles. Ultramicroscopy 53, 251–270 (1994).
Smith, S. & Abraham, S. Fatty acid synthetase from lactating rat mammary gland. 3. Dissociation and reassociation. J. Biol. Chem. 246, 6428–6435 (1971).
Montesano-Roditis, L., Glitz, D.G., Traut, R.R. & Stewart, P.L. Cryo-electron microscopic localization of protein L7/L12 within the Escherichia coli 70 S ribosome by difference mapping and Nanogold labeling. J. Biol. Chem. 276, 14117–14123 (2001).
Buchel, C., Morris, E., Orlova, E. & Barber, J. Localisation of the PsbH subunit in photosystem II: a new approach using labelling of His-tags with a Ni(2+)-NTA gold cluster and single particle analysis. J. Mol. Biol. 312, 371–379 (2001).
Witkowski, A., Joshi, A., Witkowska, H.E., Asturias, F.J. & Smith, S. Head-to-head coiled arrangement of the subunits of the animal fatty acid synthase. Chem. Biol. 11, 1667–1676 (2004).
Yuan, Z.Y. & Hammes, G.G. Fluorescence studies of chicken liver fatty acid synthase. Segmental flexibility and distance measurements. J. Biol. Chem. 261, 13643–13651 (1986).
Muesing, R.A., Lornitzo, F.A., Kumar, S. & Porter, J.W. Factors affecting the reassociation and reactivation of the half-molecular weight nonidentical subunits of pigeon liver fatty acid synthetase. J. Biol. Chem. 250, 1814–1823 (1975).
Khosla, C. Harnessing the biosynthetic potential of modular polyketide synthases. Chem. Rev. 97, 2577–2590 (1997).
Staunton, J. & Weissman, K.J. Polyketide biosynthesis: a millennium review. Nat. Prod. Rep. 18, 380–416 (2001).
Staunton, J. et al. Evidence for a double-helical structure for modular polyketide synthases. Nat. Struct. Biol. 3, 188–192 (1996).
Joshi, A.K. & Smith, S. Construction of a cDNA encoding the multifunctional animal fatty acid synthase and expression in Spodoptera frugiperda cells using baculoviral vectors. Biochem. J. 296, 143–149 (1993).
Joshi, A.K. & Smith, S. Construction, expression, and characterization of a mutated animal fatty acid synthase deficient in the dehydrase function. J. Biol. Chem. 268, 22508–22513 (1993).
Witkowski, A., Joshi, A. & Smith, S. Fatty acid synthase: in vitro complementation of inactive mutants. Biochemistry 35, 10569–10575 (1996).
Joshi, A.K., Witkowski, A. & Smith, S. Mapping of functional interactions between domains of the animal fatty acid synthase by mutant complementation in vitro. Biochemistry 36, 2316–2322 (1997).
Tischendorf, G.W., Zeichhardt, H. & Stoffler, G. Determination of the location of proteins L14, L17, L18, L19, L22, L23 on the surface of the 5oS ribosomal subunit of Escherichia coli by immune electron microscopy. Mol. Gen. Genet. 134, 187–208 (1974).
Stoffler, G. & Stoffler-Meilicke, M. The ultrastructure of macromolecular complexes studied with antibodies. In Modern Methods in Protein Chemistry (ed. Tesche, H.) 409–455 (De Gruyter, Berlin, 1983).
Frank, J. et al. SPIDER and WEB: Processing and visualization of images in 3D electron microscopy and related fields. J. Struct. Biol. 116, 190–199 (1996).
Witkowski, A., Joshi, A.K., Lindqvist, Y. & Smith, S. Conversion of a β-ketoacyl synthase to a malonyl decarboxylase by replacement of the active-site cysteine with glutamine. Biochemistry 38, 11643–11650 (1999).
Witkowski, A., Joshi, A.K. & Smith, S. Characterization of the β-carbon processing reactions of the mammalian cytosolic fatty acid synthase: role of the central core. Biochemistry 43, 10458–10466 (2004).
Witkowski, A., Joshi, A.K. & Smith, S. Mechanism of the β-ketoacyl synthase reaction catalyzed by the animal fatty acid synthase. Biochemistry 41, 10877–10887 (2002).
Acknowledgements
F.J.A. is a scholar of the Leukemia and Lymphoma Society of America. Work in the laboratory of S.S. is supported by US National Institutes of Health grant DK16073.
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Supplementary information
Supplementary Fig. 1
Raw images of FAS particles. (PDF 715 kb)
Supplementary Fig. 2
Characteristics of final ice-FAS reconstruction from unstained specimens. (PDF 96 kb)
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Asturias, F., Chadick, J., Cheung, I. et al. Structure and molecular organization of mammalian fatty acid synthase. Nat Struct Mol Biol 12, 225–232 (2005). https://doi.org/10.1038/nsmb899
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DOI: https://doi.org/10.1038/nsmb899
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