title: Protein S-glutathionylation in malaria parasites.
abstract: OBJECTIVE
  Protein S-glutathionylation is a widely distributed post-translational modification of thiol groups with glutathione that can function as a redox-sensitive switch to mediate redox regulation and signal transduction. The malaria parasite Plasmodium falciparum is exposed to intense oxidative stress and possesses the enzymatic system required to regulate protein S-glutathionylation, but despite its potential importance, protein S-glutathionylation has not yet been studied in malaria parasites. In this work we applied a method based on enzymatic deglutathionylation, affinity purification of biotin-maleimide-tagged proteins, and proteomic analyses to characterize the Plasmodium glutathionylome.
  
  
  RESULTS
  We identified 493 targets of protein S-glutathionylation in Plasmodium. Functional profiles revealed that the targets are components of central metabolic pathways, such as nitrogen compound metabolism and protein metabolism. Fifteen identified proteins with important functions in metabolic pathways (thioredoxin reductase, thioredoxin, thioredoxin peroxidase 1, glutathione reductase, glutathione S-transferase, plasmoredoxin, mitochondrial dihydrolipoamide dehydrogenase, glutamate dehydrogenase 1, glyoxalase I and II, ornithine δ-aminotransferase, lactate dehydrogenase, glyceraldehyde 3-phosphate dehydrogenase [GAPDH], pyruvate kinase [PK], and phosphoglycerate mutase) were further analyzed to study their ability to form mixed disulfides with glutathione. We demonstrate that P. falciparum GAPDH, PK, and ornithine δ-aminotransferase are reversibly inhibited by S-glutathionylation. Further, we provide evidence that not only P. falciparum glutaredoxin 1, but also thioredoxin 1 and plasmoredoxin are able to efficiently catalyze protein deglutathionylation.
  
  
  METHODS
  We used an affinity-purification based proteomic approach to characterize the Plasmodium glutathionylome.
  
  
  CONCLUSIONS
  Our results indicate a wide regulative use of S-glutathionylation in the malaria parasite and contribute to our understanding of redox-regulatory processes in this pathogen.
authors: Interdisciplinary Research Center, Justus Liebig University, Giessen, Germany.
authors: Sebastian
authors: S
authors: Kehr
authors: Esther
authors: E
authors: Jortzik
authors: Claire
authors: C
authors: Delahunty
authors: John R
authors: JR
authors: Yates
authors: Stefan
authors: S
authors: Rahlfs
authors: Katja
authors: K
authors: Becker
references: N Engl J Med. 2008 Apr 24;358(17):1805-10
references: 18420493
references: Biochem Biophys Res Commun. 1998 Jun 18;247(2):481-6
references: 9642155
references: Annu Rev Biochem. 1983;52:711-60
references: 6137189
references: Biochemistry. 2008 Jan 8;47(1):473-8
references: 18081316
references: Curr Protoc Bioinformatics. 2007 Jan;Chapter 13:Unit 13.4
references: 18428785
references: Methods Enzymol. 2002;347:370-81
references: 11898428
references: PLoS Pathog. 2008 Nov;4(11):e1000214
references: 19023412
references: Biol Pharm Bull. 2008 Oct;31(10):1875-81
references: 18827347
references: Eur J Biochem. 2001 Mar;268(5):1404-9
references: 11231293
references: Proc Natl Acad Sci U S A. 2002 Mar 19;99(6):3505-10
references: 11904414
references: Mol Biochem Parasitol. 2010 Jan;169(1):50-4
references: 19748525
references: Physiol Plant. 2008 Jun;133(2):211-28
references: 18298409
references: Biochem J. 2011 Jun 15;436(3):641-50
references: 21443518
references: Mol Plant. 2009 Mar;2(2):218-35
references: 19825609
references: J Biol Chem. 2008 Aug 1;283(31):21571-8
references: 18534986
references: Biochem J. 2003 Sep 1;374(Pt 2):513-9
references: 12755685
references: Nature. 2010 Dec 23;468(7327):1115-8
references: 21179168
references: Nat Cell Biol. 2005 Jul;7(7):665-74
references: 15951807
references: Biochem Biophys Res Commun. 2005 Jan 7;326(1):188-96
references: 15567170
references: PLoS One. 2010 Apr 29;5(4):e10399
references: 20454679
references: Free Radic Biol Med. 2007 Sep 15;43(6):883-98
references: 17697933
references: FEBS J. 2007 Jan;274(1):212-26
references: 17140414
references: Ann N Y Acad Sci. 2002 Nov;973:488-504
references: 12485918
references: Biochim Biophys Acta. 2006 Mar;1760(3):380-7
references: 16515838
references: J Biol Chem. 2009 Jan 2;284(1):436-445
references: 18990698
references: Bioinformatics. 2004 Aug 4;20 Suppl 1:i49-54
references: 15262780
references: J Biol Chem. 2009 Dec 25;284(52):36282-36291
references: 19847013
references: Antioxid Redox Signal. 2005 Mar-Apr;7(3-4):348-66
references: 15706083
references: Science. 2009 Aug 14;325(5942):834-40
references: 19608861
references: Proc Natl Acad Sci U S A. 2005 Feb 1;102(5):1357-9
references: 15677321
references: BMC Biochem. 2010 Jan 14;11:3
references: 20074363
references: Cell. 2003 Jul 25;114(2):255-66
references: 12887926
references: Biochem Biophys Res Commun. 2005 Jul 1;332(2):362-9
references: 15910747
references: J Biol Chem. 2000 Aug 25;275(34):26556-65
references: 10854441
references: FEBS Lett. 2010 Jun 3;584(11):2242-8
references: 20406640
references: J Proteome Res. 2002 May-Jun;1(3):211-5
references: 12645897
references: Arch Biochem Biophys. 2002 Oct 15;406(2):229-40
references: 12361711
references: J Biol Chem. 2001 Oct 5;276(40):37133-40
references: 11479312
references: FEBS Lett. 1998 Feb 6;422(3):311-4
references: 9498806
references: PLoS Pathog. 2009 Apr;5(4):e1000383
references: 19360125
references: J Proteome Res. 2002 Jan-Feb;1(1):21-6
references: 12643522
references: Antioxid Redox Signal. 2009 May;11(5):1059-81
references: 19119916
references: Biol Chem. 2002 May;383(5):821-30
references: 12108547
references: Mol Cell Proteomics. 2006 Feb;5(2):215-25
references: 16223748
references: J Proteome Res. 2003 Jan-Feb;2(1):43-50
references: 12643542
references: J Mol Biol. 2010 Sep 17;402(2):445-59
references: 20673832
references: FASEB J. 2006 Mar;20(3):518-20
references: 16415107
references: Curr Protoc Bioinformatics. 2010 Jun;Chapter 13:Unit 13.13.1-15
references: 20521246
references: Bosn J Basic Med Sci. 2009 Nov;9(4):278-82
references: 20001992