title: Thiol redox sensitivity of two key enzymes of heme biosynthesis and pentose phosphate pathways: uroporphyrinogen decarboxylase and transketolase.
abstract: Uroporphyrinogen decarboxylase (Hem12p) and transketolase (Tkl1p) are key mediators of two critical processes within the cell, heme biosynthesis, and the nonoxidative part of the pentose phosphate pathway (PPP). The redox properties of both Hem12p and Tkl1p from Saccharomyces cerevisiae were investigated using proteomic techniques (SRM and label-free quantification) and biochemical assays in cell extracts and in vitro with recombinant proteins. The in vivo analysis revealed an increase in oxidized Cys-peptides in the absence of Grx2p, and also after treatment with H2O2 in the case of Tkl1p, without corresponding changes in total protein, demonstrating a true redox response. Out of three detectable Cys residues in Hem12p, only the conserved residue Cys52 could be modified by glutathione and efficiently deglutathionylated by Grx2p, suggesting a possible redox control mechanism for heme biosynthesis. On the other hand, Tkl1p activity was sensitive to thiol redox modification and although Cys622 could be glutathionylated to a limited extent, it was not a natural substrate of Grx2p. The human orthologues of both enzymes have been involved in certain cancers and possess Cys residues equivalent to those identified as redox sensitive in yeast. The possible implication for redox regulation in the context of tumour progression is put forward.
authors: Department of Biochemistry and Molecular Biology, University of Córdoba and Córdoba Maimónides Institute for Biomedical Research (IMIBIC), 14071 Córdoba, Spain.
authors: Brian
authors: B
authors: McDonagh
authors: José Rafael
authors: JR
authors: Pedrajas
authors: C Alicia
authors: CA
authors: Padilla
authors: José Antonio
authors: JA
authors: Bárcena
mesh_terms: D006418:Heme / Q000096:biosynthesis*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D010427:Pentose Phosphate Pathway / Q000187:drug effects*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000378:metabolism*; D014174:Transketolase / Q000378:metabolism*; D014575:Uroporphyrinogen Decarboxylase / Q000378:metabolism*
chemical_list: D013438:Sulfhydryl Compounds; D006418:Heme; D006861:Hydrogen Peroxide; D014174:Transketolase; D014575:Uroporphyrinogen Decarboxylase
references: Hum Mol Genet. 2001 Oct 1;10(20):2181-6
references: 11673399
references: Proc Natl Acad Sci U S A. 2003 Jan 7;100(1):370-5
references: 12509500
references: FEBS J. 2007 Jan;274(1):212-26
references: 17140414
references: Plant Physiol Biochem. 2011 Sep;49(9):946-57
references: 21782461
references: J Mol Biol. 1994 May 6;238(3):387-404
references: 8176731
references: Antioxid Redox Signal. 2011 Jun 15;14(12):2581-642
references: 21275844
references: Antioxid Redox Signal. 2012 Dec 15;17(12):1748-63
references: 22530666
references: Proteomics. 2011 Feb;11(4):535-53
references: 21243637
references: Eur J Biochem. 1990 Mar 10;188(2):393-403
references: 2180703
references: Cell Mol Life Sci. 1999 Oct 30;56(5-6):415-26
references: 11212295
references: EMBO J. 1998 May 1;17(9):2463-71
references: 9564029
references: Cell. 2011 Mar 4;144(5):646-74
references: 21376230
references: Free Radic Biol Med. 2007 Jul 1;43(1):62-70
references: 17561094
references: Arch Biochem Biophys. 1959 May;82(1):70-7
references: 13650640
references: J Cell Physiol. 2012 Feb;227(2):421-30
references: 21448925
references: Electrophoresis. 1997 Dec;18(15):2714-23
references: 9504803
references: Am J Physiol Cell Physiol. 2008 Oct;295(4):C849-68
references: 18684987
references: J Biol Chem. 2009 Aug 28;284(35):23364-74
references: 19561357
references: Mol Biol Cell. 2002 Apr;13(4):1109-21
references: 11950925
references: Biochem J. 2004 Sep 1;382(Pt 2):759-67
references: 15149284
references: Phytochemistry. 2010 Jun;71(8-9):853-9
references: 20417532
references: Biosci Rep. 2004 Aug-Oct;24(4-5):523-44
references: 16134024
references: Mol Cell Biol. 1999 Apr;19(4):2650-6
references: 10082531
references: J Proteomics. 2011 Oct 19;74(11):2487-97
references: 21565288
references: Br J Cancer. 2006 Feb 27;94(4):578-85
references: 16465194
references: Biochim Biophys Acta. 2009 Dec;1796(2):252-65
references: 19682552
references: Science. 2011 Dec 2;334(6060):1278-83
references: 22052977
references: Arch Biochem Biophys. 2008 Jun 15;474(2):238-51
references: 18314007
references: Eur J Biochem. 2001 Aug;268(15):4177-82
references: 11488910
references: Sci Transl Med. 2011 Jan 26;3(67):67ra7
references: 21270338
references: EMBO J. 1992 Jul;11(7):2373-9
references: 1628611
references: J Biol Chem. 2004 Mar 26;279(13):12386-95
references: 14722110
references: Cell Metab. 2013 Mar 5;17(3):319-28
references: 23473029
references: J Bioenerg Biomembr. 1995 Apr;27(2):207-14
references: 7592567
references: Eur J Biochem. 1994 May 1;221(3):1033-7
references: 8181459
references: Antioxid Redox Signal. 2011 Oct 15;15(8):2335-81
references: 21194351
references: Nat Rev Cancer. 2010 Apr;10(4):267-77
references: 20300106
references: Biochem J. 1929;23(3):536-45
references: 16744238
references: J Proteomics. 2009 May 2;72(4):677-89
references: 19367685
references: J Cell Sci. 2006 Nov 1;119(Pt 21):4554-64
references: 17074835
references: Biochem J. 2002 Jun 15;364(Pt 3):617-23
references: 11958675
references: Antioxid Redox Signal. 2011 Jul 1;15(1):135-46
references: 20812876
references: Antioxid Redox Signal. 2011 Mar 15;14(6):1065-77
references: 20799881
references: Biochem Biophys Res Commun. 1966 Jan 4;22(1):129-34
references: 5937329
references: J Biol Chem. 2011 Apr 29;286(17):15565-76
references: 21385868
references: Antioxid Redox Signal. 2008 Nov;10(11):1941-88
references: 18774901
references: J Biol Chem. 2004 Jul 9;279(28):29101-8
references: 15128732
references: Arch Biochem Biophys. 2004 Jun 1;426(1):43-54
references: 15130781
references: Nat Methods. 2008 Nov;5(11):913-4
references: 18974732
references: Open Biol. 2013 Jan 08;3(1):120144
references: 23303309
references: J Biol Chem. 1994 Oct 7;269(40):25010-5
references: 7929187
references: Science. 1956 Feb 24;123(3191):309-14
references: 13298683