title: The Succinated Proteome of FH-Mutant Tumours.
abstract: Inherited mutations in the Krebs cycle enzyme fumarate hydratase (FH) predispose to hereditary leiomyomatosis and renal cell cancer (HLRCC). Loss of FH activity in HLRCC tumours causes accumulation of the Krebs cycle intermediate fumarate to high levels, which may act as an oncometabolite through various, but not necessarily mutually exclusive, mechanisms. One such mechanism, succination, is an irreversible non-enzymatic modification of cysteine residues by fumarate, to form S-(2-succino)cysteine (2SC). Previous studies have demonstrated that succination of proteins including glyceraldehyde 3-phosphate dehydrogenase (GAPDH), kelch-like ECH-associated protein 1 (KEAP1) and mitochondrial aconitase (ACO2) can have profound effects on cellular metabolism. Furthermore, immunostaining for 2SC is a sensitive and specific biomarker for HLRCC tumours. Here, we performed a proteomic screen on an FH-mutant tumour and two HLRCC-derived cancer cell lines and identified 60 proteins where one or more cysteine residues were succinated; 10 of which were succinated at cysteine residues either predicted, or experimentally proven, to be functionally significant. Bioinformatic enrichment analyses identified most succinated targets to be involved in redox signaling. To our knowledge, this is the first proteomic-based succination screen performed in human tumours and cancer-derived cells and has identified novel 2SC targets that may be relevant to the pathogenesis of HLRCC.
authors.affiliation: Cancer Biology and Metabolism Group, Institute of Genetics and Molecular Medicine, Edinburgh Cancer Research UK Centre, University of Edinburgh, Western General Hospital, Crewe Road South, Edinburgh EH4 2XR, UK. ming.yang@igmm.ed.ac.uk.
authors.forename: Ming
authors.initials: M
authors.lastname: Yang
authors.affiliation: Central Proteomics Facility, Target Discovery Institute, University of Oxford, Oxford OX3 7FZ, UK. nicola.ternette@ndm.ox.ac.uk.
authors.forename: Nicola
authors.initials: N
authors.lastname: Ternette
authors.affiliation: Cancer Biology and Metabolism Group, Institute of Genetics and Molecular Medicine, Edinburgh Cancer Research UK Centre, University of Edinburgh, Western General Hospital, Crewe Road South, Edinburgh EH4 2XR, UK. huizhong.su@igmm.ed.ac.uk.
authors.forename: Huizhong
authors.initials: H
authors.lastname: Su
authors.affiliation: Cancer Biology and Metabolism Group, Institute of Genetics and Molecular Medicine, Edinburgh Cancer Research UK Centre, University of Edinburgh, Western General Hospital, Crewe Road South, Edinburgh EH4 2XR, UK. rd292@exeter.ac.uk.
authors.forename: Raliat
authors.initials: R
authors.lastname: Dabiri
authors.affiliation: Central Proteomics Facility, Target Discovery Institute, University of Oxford, Oxford OX3 7FZ, UK. benedikt.kessler@ndm.ox.ac.uk.
authors.forename: Benedikt M
authors.initials: BM
authors.lastname: Kessler
authors.affiliation: Oxford Centre for Diabetes, Endocrinology & Metabolism, University of Oxford, Oxford OX3 7LJ, UK. julie.adam@ocdem.ox.ac.uk.
authors.forename: Julie
authors.initials: J
authors.lastname: Adam
authors.affiliation: Cancer Science Institute of Singapore, National University of Singapore, Centre for Translational Medicine, #12-01,117599, Singapore. teh.bin.tean@singhealth.com.sg.
authors.forename: Bin Tean
authors.initials: BT
authors.lastname: Teh
authors.affiliation: Cancer Biology and Metabolism Group, Institute of Genetics and Molecular Medicine, Edinburgh Cancer Research UK Centre, University of Edinburgh, Western General Hospital, Crewe Road South, Edinburgh EH4 2XR, UK. patrick.pollard@igmm.ed.ac.uk.
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Pollard
references.citation: Mass Spectrom Rev. 2014 Mar-Apr;33(2):98-109
references.pmid: 24115015
references.citation: J Lipid Res. 2011 Feb;52(2):393-8
references.pmid: 21044946
references.citation: Hum Mol Genet. 2013 Dec 1;22(23):4805-17
references.pmid: 23847046
references.citation: Nat Protoc. 2009;4(1):44-57
references.pmid: 19131956
references.citation: J Lab Clin Med. 1997 Mar;129(3):318-29
references.pmid: 9042817
references.citation: Free Radic Res. 2011 Jan;45(1):101-9
references.pmid: 20964553
references.citation: Cell Rep. 2013 May 30;3(5):1440-8
references.pmid: 23643539
references.citation: Proc Natl Acad Sci U S A. 2009 Jan 27;106(4):1111-6
references.pmid: 19144925
references.citation: Biochemistry. 1996 May 28;35(21):6735-44
references.pmid: 8639624
references.citation: Mol Cell. 2014 Feb 20;53(4):645-54
references.pmid: 24486019
references.citation: Diabetes. 2008 Jan;57(1):41-9
references.pmid: 17934141
references.citation: Cell Rep. 2013 Mar 28;3(3):689-700
references.pmid: 23499446
references.citation: J Pathol. 2011 Sep;225(1):4-11
references.pmid: 21630274
references.citation: Biochim Biophys Acta. 2014 Feb;1840(2):906-12
references.pmid: 23939310
references.citation: Proc Natl Acad Sci U S A. 2007 Jul 10;104(28):11609-14
references.pmid: 17606900
references.citation: Proc Natl Acad Sci U S A. 2014 Mar 18;111(11):4025-30
references.pmid: 24550463
references.citation: Electrophoresis. 1999 Dec;20(18):3551-67
references.pmid: 10612281
references.citation: Oxid Med Cell Longev. 2013;2013:836760
references.pmid: 23983902
references.citation: Nat Rev Neurosci. 2006 Mar;7(3):207-19
references.pmid: 16495942
references.citation: J Mol Cell Cardiol. 2014 Aug;73:2-9
references.pmid: 24512843
references.citation: Sci Transl Med. 2012 Mar 28;4(127):127ps10
references.pmid: 22461637
references.citation: Oncogene. 2014 May 15;33(20):2547-56
references.pmid: 23812428
references.citation: Kidney Int. 2007 Jul;72(2):157-65
references.pmid: 17396115
references.citation: Proc Natl Acad Sci U S A. 2010 May 18;107(20):9117-22
references.pmid: 20439756
references.citation: Proc Natl Acad Sci U S A. 2014 Mar 18;111(11):4019-24
references.pmid: 24550462
references.citation: Proc Natl Acad Sci U S A. 2014 Mar 18;111(11):4031-6
references.pmid: 24550447
references.citation: Hum Pathol. 2012 Apr;43(4):577-84
references.pmid: 21943684
references.citation: Cancer Res. 2013 Apr 1;73(7):2044-51
references.pmid: 23365135
references.citation: PLoS Biol. 2004 Nov;2(11):e362
references.pmid: 15502874
references.citation: Biochem Biophys Res Commun. 2003 Apr 25;304(1):176-83
references.pmid: 12705903
references.citation: Proc Natl Acad Sci U S A. 2004 Jun 15;101(24):9103-8
references.pmid: 15181200
references.citation: Free Radic Res. 2006 Feb;40(2):155-65
references.pmid: 16390825
references.citation: Nucleic Acids Res. 2007 Jul;35(Web Server issue):W169-75
references.pmid: 17576678
references.citation: Trends Biochem Sci. 2003 Jan;28(1):32-40
references.pmid: 12517450
references.citation: Nat Chem Biol. 2005 Aug;1(3):154-8
references.pmid: 16408020
references.citation: J Clin Invest. 2013 Sep;123(9):3652-8
references.pmid: 23999438
references.citation: Biochim Biophys Acta. 1993 Aug 7;1164(3):268-72
references.pmid: 8343525
references.citation: Cancer Cell. 2011 Oct 18;20(4):524-37
references.pmid: 22014577
references.citation: Front Oncol. 2012 May 18;2:49
references.pmid: 22655275
references.citation: J Biol Chem. 2011 Feb 11;286(6):4098-106
references.pmid: 21097510
references.citation: Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3633-8
references.pmid: 9108029
references.citation: J Biol Chem. 2007 Nov 23;282(47):34219-28
references.pmid: 17726021
references.citation: Bioinformatics. 2010 Apr 15;26(8):1131-2
references.pmid: 20189941
references.citation: Mol Cell. 2013 Jul 25;51(2):236-48
references.pmid: 23747014
references.citation: Cancer Genet Cytogenet. 2010 Jan 1;196(1):45-55
references.pmid: 19963135
references.citation: Free Radic Biol Med. 2010 Nov 1;49(8):1283-9
references.pmid: 20673799
references.citation: Trends Endocrinol Metab. 2012 Nov;23(11):552-9
references.pmid: 22858391
references.citation: Biochem J. 2012 Jul 15;445(2):247-54
references.pmid: 22524437
references.citation: Nat Chem Biol. 2008 Mar;4(3):152-6
references.pmid: 18277970
references.citation: Cancer Cell. 2011 Oct 18;20(4):511-23
references.pmid: 22014576
references.citation: Front Oncol. 2012 Jul 31;2:85
references.pmid: 22866264
references.citation: Obesity (Silver Spring). 2012 Feb;20(2):263-9
references.pmid: 22134201
references.citation: Arch Biochem Biophys. 2006 Jun 1;450(1):1-8
references.pmid: 16624247
references.citation: Nature. 2014 Jun 19;510(7505):422-426
references.pmid: 24814345
references.citation: Biochem J. 2014 Sep 1;462(2):231-45
references.pmid: 24909641
references.citation: Hum Mol Genet. 2010 Jun 15;19(12):2395-408
references.pmid: 20304780
references.citation: J Neurosci Res. 2009 Jan;87(1):123-9
references.pmid: 18711745
references.citation: Dis Model Mech. 2012 Mar;5(2):155-64
references.pmid: 22382365
references.citation: World J Urol. 2013 Oct;31(5):1191-6
references.pmid: 22544372
references.citation: Nucleic Acids Res. 2009 Jan;37(Database issue):D412-6
references.pmid: 18940858
references.citation: Hum Mol Genet. 2010 Oct 1;19(19):3844-51
references.pmid: 20660115