title: Structural consequences of cysteinylation of Cu/Zn-superoxide dismutase.
abstract: The metalloenzyme Cu/Zn-superoxide dismutase (SOD1) catalyzes the reduction of superoxide anions into molecular oxygen and hydrogen peroxide. Hydrogen peroxide can oxidize SOD1, resulting in aberrant protein conformational changes, disruption of SOD1 function, and DNA damage. Cells may have evolved mechanisms of regulation that prevent such oxidation. We observed that cysteinylation of cysteine 111 (Cys111) of SOD1 prevents oxidation by peroxide (DOI 10.1021/bi4006122 ). In this article, we characterize cysteinylated SOD1 using differential scanning fluorometry and X-ray crystallography. The stoichiometry of binding was one cysteine per SOD1 dimer, and there does not appear to be free volume for a second cysteine without disrupting the dimer interface. Much of the three-dimensional structure of SOD1 is unaffected by cysteinylation. However, local conformational changes are observed in the cysteinylated monomer that include changes in conformation of the electrostatic loop (loop VII; residues 133-144) and the dimer interface (loop VI; residues 102-115). In addition, our data shows how cysteinylation precludes oxidation of cysteine 111 and suggests possible cross-talk between the dimer interface and the electrostatic loop.
authors.affiliation: Departments of Biochemistry and Chemistry and Rosenstiel Basic Medical Sciences Research Center, Brandeis University , Waltham, Massachusetts 02454, United States.
authors.forename: Jared R
authors.initials: JR
authors.lastname: Auclair
authors.forename: Heather R
authors.initials: HR
authors.lastname: Brodkin
authors.forename: J Alejandro
authors.initials: JA
authors.lastname: D'Aquino
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Petsko
authors.forename: Dagmar
authors.initials: D
authors.lastname: Ringe
authors.forename: Jeffrey N
authors.initials: JN
authors.lastname: Agar
mesh_terms: D018360:Crystallography, X-Ray; D003545:Cysteine / Q000737:chemistry*; D004795:Enzyme Stability / Q000187:drug effects; D005470:Fluorometry; D008958:Models, Molecular; D010084:Oxidation-Reduction; D011487:Protein Conformation / Q000187:drug effects; D055503:Protein Multimerization; D011499:Protein Processing, Post-Translational; D013482:Superoxide Dismutase / Q000378:metabolism*; D000072105:Superoxide Dismutase-1
chemical_list: D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D003545:Cysteine
references.citation: J Biol Chem. 2004 Jul 30;279(31):32534-40
references.pmid: 15123612
references.citation: Nat Struct Biol. 2003 Jun;10(6):461-7
references.pmid: 12754496
references.citation: J Biol Chem. 2002 May 3;277(18):15923-31
references.pmid: 11854284
references.citation: J Biol Chem. 1987 Jun 15;262(17):8227-34
references.pmid: 3597373
references.citation: J Biol Chem. 2007 Sep 7;282(36):25981-5
references.pmid: 17611193
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
references.pmid: 15572765
references.citation: Acta Crystallogr D Biol Crystallogr. 2002 Nov;58(Pt 11):1948-54
references.pmid: 12393927
references.citation: J Mol Biol. 2007 Jan 12;365(2):333-42
references.pmid: 17070542
references.citation: J Biol Chem. 2004 Apr 9;279(15):15499-504
references.pmid: 14734542
references.citation: J Biol Chem. 1968 Nov 10;243(21):5753-60
references.pmid: 4972775
references.citation: Acta Crystallogr D Biol Crystallogr. 2003 Jul;59(Pt 7):1124-30
references.pmid: 12832754
references.citation: Proc Natl Acad Sci U S A. 1995 Sep 12;92(19):8574-8
references.pmid: 7567977
references.citation: J Biol Chem. 2007 Jun 1;282(22):16329-35
references.pmid: 17389599
references.citation: Nat Med. 2007 Jun;13(6):754-9
references.pmid: 17486090
references.citation: Genome. 2004 Dec;47(6):1029-35
references.pmid: 15644960
references.citation: Ann Neurol. 2007 Dec;62(6):553-9
references.pmid: 18074357
references.citation: Hear Res. 2005 Nov;209(1-2):76-85
references.pmid: 16055286
references.citation: J Biol Chem. 2004 Dec 24;279(52):54558-66
references.pmid: 15485869
references.citation: Biosci Rep. 2012 Dec;32(6):539-48
references.pmid: 22804629
references.citation: Proc Natl Acad Sci U S A. 2010 Dec 14;107(50):21394-9
references.pmid: 21098299
references.citation: Methods Enzymol. 1997;276:307-26
references.pmid: 27754618
references.citation: J Biol Chem. 2003 Jun 27;278(26):24078-89
references.pmid: 12686560
references.citation: Neurosci Lett. 1994 Sep 26;179(1-2):149-52
references.pmid: 7845611
references.citation: Anal Biochem. 2001 Aug 1;295(1):45-56
references.pmid: 11476544
references.citation: Science. 1993 Aug 20;261(5124):1047-51
references.pmid: 8351519
references.citation: J Biol Chem. 2009 Nov 6;284(45):30965-73
references.pmid: 19635794
references.citation: Biochemistry. 2013 Sep 10;52(36):6137-44
references.pmid: 23927036
references.citation: J Biol Chem. 2005 Mar 25;280(12):11648-55
references.pmid: 15659387
references.citation: J Biol Chem. 1987 Jun 15;262(17):8220-6
references.pmid: 3597372
references.citation: J Biol Chem. 2002 Dec 6;277(49):47551-6
references.pmid: 12356748
references.citation: Proc Natl Acad Sci U S A. 2006 Oct 24;103(43):15835-40
references.pmid: 17035505
references.citation: J Biol Chem. 2007 Dec 7;282(49):35933-44
references.pmid: 17913710
references.citation: Ann Neurol. 1997 Aug;42(2):135-7
references.pmid: 9266721
references.citation: Proc Natl Acad Sci U S A. 2006 May 23;103(21):8060-5
references.pmid: 16690741
references.citation: Rapid Commun Mass Spectrom. 2005;19(20):2965-73
references.pmid: 16178042