title: Inhibition of mitochondrial aconitase by succination in fumarate hydratase deficiency.
abstract: The gene encoding the Krebs cycle enzyme fumarate hydratase (FH) is mutated in hereditary leiomyomatosis and renal cell cancer (HLRCC). Loss of FH activity causes accumulation of intracellular fumarate, which can directly modify cysteine residues to form 2-succinocysteine through succination. We undertook a proteomic-based screen in cells and renal cysts from Fh1 (murine FH)-deficient mice and identified 94 protein succination targets. Notably, we identified the succination of three cysteine residues in mitochondrial Aconitase2 (ACO2) crucial for iron-sulfur cluster binding. We show that fumarate exerts a dose-dependent inhibition of ACO2 activity, which correlates with increased succination as determined by mass spectrometry, possibly by interfering with iron chelation. Importantly, we show that aconitase activity is impaired in FH-deficient cells. Our data provide evidence that succination, resulting from FH deficiency, targets and potentially alters the function of multiple proteins and may contribute to the dysregulated metabolism observed in HLRCC.
authors.affiliation: Central Proteomics Facility, Henry Wellcome Building for Molecular Physiology, University of Oxford, Oxford OX3 7BN, UK.
authors.forename: Nicola
authors.initials: N
authors.lastname: Ternette
authors.forename: Ming
authors.initials: M
authors.lastname: Yang
authors.forename: Mahima
authors.initials: M
authors.lastname: Laroyia
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Kitagawa
authors.forename: Linda
authors.initials: L
authors.lastname: O'Flaherty
authors.forename: Kathryn
authors.initials: K
authors.lastname: Wolhulter
authors.forename: Kaori
authors.initials: K
authors.lastname: Igarashi
authors.forename: Kaori
authors.initials: K
authors.lastname: Saito
authors.forename: Keiko
authors.initials: K
authors.lastname: Kato
authors.forename: Roman
authors.initials: R
authors.lastname: Fischer
authors.forename: Alexandre
authors.initials: A
authors.lastname: Berquand
authors.forename: Benedikt M
authors.initials: BM
authors.lastname: Kessler
authors.forename: Terry
authors.initials: T
authors.lastname: Lappin
authors.forename: Norma
authors.initials: N
authors.lastname: Frizzell
authors.forename: Tomoyoshi
authors.initials: T
authors.lastname: Soga
authors.forename: Julie
authors.initials: J
authors.lastname: Adam
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Pollard
mesh_terms: D000154:Aconitate Hydratase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D003545:Cysteine / Q000378:metabolism; D005649:Fumarate Hydratase / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism*; D005650:Fumarates / Q000378:metabolism; D006801:Humans; D007501:Iron / Q000378:metabolism; D007680:Kidney Neoplasms / Q000378:metabolism*; D018231:Leiomyomatosis / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D008928:Mitochondria / Q000378:metabolism*; D009386:Neoplastic Syndromes, Hereditary / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism; D012878:Skin Neoplasms; D019802:Succinic Acid / Q000378:metabolism*; D014594:Uterine Neoplasms
chemical_list: D005650:Fumarates; D020543:Proteome; C032005:fumaric acid; D019802:Succinic Acid; D007501:Iron; D005649:Fumarate Hydratase; C476079:ACO2 protein, human; D000154:Aconitate Hydratase; D003545:Cysteine