Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
BioCyc websites down
12/28 - 12/31
for maintenance.
Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
BioCyc websites down
12/28 - 12/31
for maintenance.
Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
BioCyc websites down
12/28 - 12/31
for maintenance.
Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
BioCyc websites down
12/28 - 12/31
for maintenance.
Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
BioCyc websites down
12/28 - 12/31
for maintenance.
twitter

Escherichia coli K-12 substr. MG1655 Enzyme: 3'-phospho-adenylylsulfate reductase



Gene: cysH Accession Numbers: EG10189 (EcoCyc), b2762, ECK2757

Regulation Summary Diagram: ?

Subunit composition of 3'-phospho-adenylylsulfate reductase = [CysH]2

Summary:
3'-Phospho-adenylylsulfate reductase (CysH)is involved in the assimilation of sulfate and catalyzes the reduction of 3'-phospho-adenylylsulfate (PAPS) to sulfite. PAPS reductase is composed of two identical subunits of 28 kDa. It has no chromophores and contains a single cysteine per subunit in a highly conserved ECGLH motif, which is identified as the redox-active center of the enzyme [Berendt95, Krone91, Savage97].

Reduction of sulfate to sulfite by PAPS reductase requires two electrons which are donated by the cysteine residues of the enzyme in a ping-pong mechanism. During the transfer the cysteines are oxidized and form a disulfide. The oxidized enzyme is inactive and needs to be reduced for the reduction of PAPS to continue. Dimeric oxidized PAPS reductase migrates on SDS-PAGE with an apparent molecular mass (60 kDa) higher than that of the reduced form (30 kDa). The enzyme can be reduced in vitro by the thioredoxins Trx1 and Trx2 and the glutaredoxin Grx1 [Lillig99]. The likely in vivo relevant factor is Trx1.

The enzyme is also regulated by glutathione: if the intracellular environment is somewhat oxidizing, a mixed disulfide may form between Cys239 of the enzyme and glutathione, rendering the enzyme inactive. This inactivation can be relieved by all glutaredoxins, which can catalyze the reduction of this mixed disulfide [Lillig03].

A conserved residue which altered catalytic activity when mutated was identified through site directed mutagenesis studies [Berendt95].

The crystal structure of CysH in complex with thioredoxin I has been determined at 3.0 Å resolution [Chartron07].

Gene Citations: [Ostrowski89]

Locations: cytosol

Map Position: [2,885,600 <- 2,886,334] (62.19 centisomes)
Length: 735 bp / 244 aa

Molecular Weight of Polypeptide: 27.976 kD (from nucleotide sequence), 30.0 kD (experimental) [Lillig99 ]

Molecular Weight of Multimer: 60.0 kD (experimental) [Lillig99]

Unification Links: ASAP:ABE-0009057 , CGSC:892 , EchoBASE:EB0186 , EcoGene:EG10189 , EcoliWiki:b2762 , ModBase:P17854 , OU-Microarray:b2762 , PortEco:cysH , PR:PRO_000022379 , Pride:P17854 , Protein Model Portal:P17854 , RefSeq:NP_417242 , RegulonDB:EG10189 , SMR:P17854 , String:511145.b2762 , UniProt:P17854

Relationship Links: InterPro:IN-FAMILY:IPR002500 , InterPro:IN-FAMILY:IPR004511 , InterPro:IN-FAMILY:IPR011800 , InterPro:IN-FAMILY:IPR014729 , PDB:Structure:1SUR , PDB:Structure:2O8V , Pfam:IN-FAMILY:PF01507

In Paralogous Gene Group: 452 (2 members)

Gene-Reaction Schematic: ?

Genetic Regulation Schematic: ?

GO Terms:

Biological Process: GO:0006790 - sulfur compound metabolic process Inferred from experiment [Berendt95]
GO:0019379 - sulfate assimilation, phosphoadenylyl sulfate reduction by phosphoadenylyl-sulfate reductase (thioredoxin) Inferred from experiment Inferred by computational analysis [GOA06, GOA01, Berendt95]
GO:0008152 - metabolic process Inferred by computational analysis [GOA01]
GO:0009086 - methionine biosynthetic process Inferred by computational analysis [GOA01]
GO:0019344 - cysteine biosynthetic process Inferred by computational analysis [GOA01]
GO:0055114 - oxidation-reduction process Inferred by computational analysis [UniProtGOA11, GOA01]
GO:0070814 - hydrogen sulfide biosynthetic process Inferred by computational analysis [UniProtGOA12]
Molecular Function: GO:0004604 - phosphoadenylyl-sulfate reductase (thioredoxin) activity Inferred from experiment Inferred by computational analysis [GOA06, GOA01a, GOA01, Berendt95]
GO:0003824 - catalytic activity Inferred by computational analysis [GOA01]
GO:0016491 - oxidoreductase activity Inferred by computational analysis [UniProtGOA11]
Cellular Component: GO:0005737 - cytoplasm Inferred by computational analysis [UniProtGOA11a, UniProtGOA11, GOA06]
GO:0005829 - cytosol Inferred by computational analysis [DiazMejia09]

MultiFun Terms: metabolism metabolism of other compounds sulfur metabolism

Essentiality data for cysH knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enriched Yes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB Lennox Yes 37 Aerobic 7   Yes [Baba06, Comment 2]
M9 medium with 0.4% glucose No 37 Aerobic 7.2 0.27 No [Patrick07, Comment 3]
M9 medium with 1% glycerol No 37 Aerobic 7.2 0.35 No [Joyce06]
MOPS medium with 0.4% glucose Indeterminate 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]
No [Feist07, Comment 4]

Credits:
Last-Curated ? 26-Aug-2013 by Kubo A , SRI International


Enzymatic reaction of: 3'-phospho-adenylylsulfate reductase

Synonyms: PAPS sulfotransferase, sulfite reductase, thioredoxin:adenosine-3'-phosphate-5'-phosphosulfate reductase, PAPS reductase, thioredoxin:3'-phosphoadenylyl sulfate reductase

EC Number: 1.8.4.8

adenosine 3',5'-bisphosphate + sulfite + an oxidized thioredoxin + 2 H+ <=> 3'-phosphoadenylyl-sulfate + a reduced thioredoxin

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the Enzyme Commission system.

The reaction is favored in the opposite direction.

Alternative Substrates for a reduced thioredoxin: reduced glutaredoxin 1 [Lillig99 ]

In Pathways: superpathway of sulfate assimilation and cysteine biosynthesis , sulfate reduction I (assimilatory)

Inhibitors (Competitive): adenosine 3',5'-bisphosphate (Kic = 4µM) [Berendt95, Comment 5]

Kinetic Parameters:

Substrate
Km (μM)
kcat (sec-1)
kcat/Km (sec-1 μM-1)
Citations
3'-phosphoadenylyl-sulfate
10.0
[Berendt95]
3'-phosphoadenylyl-sulfate
22.5
3.5
[Lillig99, BRENDA14]
a reduced thioredoxin
23.0
[Berendt95]

T(opt): 35 °C [BRENDA14, Lillig99]

pH(opt): 9.2 [BRENDA14, Lillig99]


Sequence Features

Feature Class Location Citations Comment
Cleavage-of-Initial-Methionine 1
[Krone90, UniProt11]
UniProt: Removed.
Chain 2 -> 244
[UniProt09]
UniProt: Phosphoadenosine phosphosulfate reductase;
Sequence-Conflict 27
[Ostrowski89, UniProt10]
Alternate sequence: E → Q; UniProt: (in Ref. 2; AAA23652);
Mutagenesis-Variant 209
[UniProt10]
Alternate sequence: Y → F; UniProt: Reduction in Vmax;
Sequence-Conflict 226
[Ostrowski89, UniProt10]
Alternate sequence: A → S; UniProt: (in Ref. 2; AAA23652);
Mutagenesis-Variant 239
[UniProt10]
Alternate sequence: C → S; UniProt: 5-fold reduction in Vmax;


Gene Local Context (not to scale): ?

Transcription Units:

Notes:

History:
10/20/97 Gene b2762 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10189; confirmed by SwissProt match.


References

Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554

Berendt95: Berendt U, Haverkamp T, Prior A, Schwenn JD (1995). "Reaction mechanism of thioredoxin: 3'-phospho-adenylylsulfate reductase investigated by site-directed mutagenesis." Eur J Biochem 1995;233(1);347-56. PMID: 7588765

BRENDA14: BRENDA team (2014). "Imported from BRENDA version existing on Aug 2014." http://www.brenda-enzymes.org.

Chartron07: Chartron J, Shiau C, Stout CD, Carroll KS (2007). "3'-Phosphoadenosine-5'-phosphosulfate reductase in complex with thioredoxin: a structural snapshot in the catalytic cycle." Biochemistry 46(13);3942-51. PMID: 17352498

DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114

Dyson90: Dyson HJ, Gippert GP, Case DA, Holmgren A, Wright PE (1990). "Three-dimensional solution structure of the reduced form of Escherichia coli thioredoxin determined by nuclear magnetic resonance spectroscopy." Biochemistry 1990;29(17);4129-36. PMID: 2193685

Eklund84: Eklund H, Cambillau C, Sjoberg BM, Holmgren A, Jornvall H, Hoog JO, Branden CI (1984). "Conformational and functional similarities between glutaredoxin and thioredoxins." EMBO J 1984;3(7);1443-9. PMID: 6378624

Feist07: Feist AM, Henry CS, Reed JL, Krummenacker M, Joyce AR, Karp PD, Broadbelt LJ, Hatzimanikatis V, Palsson BO (2007). "A genome-scale metabolic reconstruction for Escherichia coli K-12 MG1655 that accounts for 1260 ORFs and thermodynamic information." Mol Syst Biol 3;121. PMID: 17593909

Gerdes03: Gerdes SY, Scholle MD, Campbell JW, Balazsi G, Ravasz E, Daugherty MD, Somera AL, Kyrpides NC, Anderson I, Gelfand MS, Bhattacharya A, Kapatral V, D'Souza M, Baev MV, Grechkin Y, Mseeh F, Fonstein MY, Overbeek R, Barabasi AL, Oltvai ZN, Osterman AL (2003). "Experimental determination and system level analysis of essential genes in Escherichia coli MG1655." J Bacteriol 185(19);5673-84. PMID: 13129938

Gleason88: Gleason FK, Holmgren A (1988). "Thioredoxin and related proteins in procaryotes." FEMS Microbiol Rev 1988;4(4);271-97. PMID: 3152490

Gleason90: Gleason FK, Lim CJ, Gerami-Nejad M, Fuchs JA (1990). "Characterization of Escherichia coli thioredoxins with altered active site residues." Biochemistry 1990;29(15);3701-9. PMID: 2187529

GOA01: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

GOA01a: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

GOA06: GOA, SIB (2006). "Electronic Gene Ontology annotations created by transferring manual GO annotations between orthologous microbial proteins."

Holmgren85: Holmgren A (1985). "Thioredoxin." Annu Rev Biochem 1985;54;237-71. PMID: 3896121

Joyce06: Joyce AR, Reed JL, White A, Edwards R, Osterman A, Baba T, Mori H, Lesely SA, Palsson BO, Agarwalla S (2006). "Experimental and computational assessment of conditionally essential genes in Escherichia coli." J Bacteriol 188(23);8259-71. PMID: 17012394

Katti90: Katti SK, LeMaster DM, Eklund H (1990). "Crystal structure of thioredoxin from Escherichia coli at 1.68 A resolution." J Mol Biol 1990;212(1);167-84. PMID: 2181145

Krone90: Krone FA, Westphal G, Meyer HE, Schwenn JD (1990). "PAPS-reductase of Escherichia coli. Correlating the N-terminal amino acid sequence with the DNA of gene cys H." FEBS Lett 260(1);6-9. PMID: 2404794

Krone91: Krone FA, Westphal G, Schwenn JD (1991). "Characterisation of the gene cysH and of its product phospho-adenylylsulphate reductase from Escherichia coli." Mol Gen Genet 1991;225(2);314-9. PMID: 2005873

Lillig03: Lillig CH, Potamitou A, Schwenn JD, Vlamis-Gardikas A, Holmgren A (2003). "Redox regulation of 3'-phosphoadenylylsulfate reductase from Escherichia coli by glutathione and glutaredoxins." J Biol Chem 278(25);22325-30. PMID: 12682041

Lillig99: Lillig CH, Prior A, Schwenn JD, Aslund F, Ritz D, Vlamis-Gardikas A, Holmgren A (1999). "New thioredoxins and glutaredoxins as electron donors of 3'-phosphoadenylylsulfate reductase." J Biol Chem 1999;274(12);7695-8. PMID: 10075658

Nikkola93: Nikkola M, Gleason FK, Fuchs JA, Eklund H (1993). "Crystal structure analysis of a mutant Escherichia coli thioredoxin in which lysine 36 is replaced by glutamic acid." Biochemistry 1993;32(19);5093-8. PMID: 8098620

Ostrowski89: Ostrowski J, Wu JY, Rueger DC, Miller BE, Siegel LM, Kredich NM (1989). "Characterization of the cysJIH regions of Salmonella typhimurium and Escherichia coli B. DNA sequences of cysI and cysH and a model for the siroheme-Fe4S4 active center of sulfite reductase hemoprotein based on amino acid homology with spinach nitrite reductase." J Biol Chem 264(26);15726-37. PMID: 2670946

Patrick07: Patrick WM, Quandt EM, Swartzlander DB, Matsumura I (2007). "Multicopy suppression underpins metabolic evolvability." Mol Biol Evol 24(12);2716-22. PMID: 17884825

Savage97: Savage H, Montoya G, Svensson C, Schwenn JD, Sinning I (1997). "Crystal structure of phosphoadenylyl sulphate (PAPS) reductase: a new family of adenine nucleotide alpha hydrolases." Structure 1997;5(7);895-906. PMID: 9261082

UniProt09: UniProt Consortium (2009). "UniProt version 15.8 released on 2009-10-01 00:00:00." Database.

UniProt10: UniProt Consortium (2010). "UniProt version 2010-11 released on 2010-11-02 00:00:00." Database.

UniProt11: UniProt Consortium (2011). "UniProt version 2011-06 released on 2011-06-30 00:00:00." Database.

UniProtGOA11: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

UniProtGOA11a: UniProt-GOA (2011). "Gene Ontology annotation based on the manual assignment of UniProtKB Subcellular Location terms in UniProtKB/Swiss-Prot entries."

UniProtGOA12: UniProt-GOA (2012). "Gene Ontology annotation based on UniPathway vocabulary mapping."

Other References Related to Gene Regulation

Drazic13: Drazic A, Miura H, Peschek J, Le Y, Bach NC, Kriehuber T, Winter J (2013). "Methionine oxidation activates a transcription factor in response to oxidative stress." Proc Natl Acad Sci U S A 110(23);9493-8. PMID: 23690622

Gebendorfer12: Gebendorfer KM, Drazic A, Le Y, Gundlach J, Bepperling A, Kastenmuller A, Ganzinger KA, Braun N, Franzmann TM, Winter J (2012). "Identification of a Hypochlorite-specific Transcription Factor from Escherichia coli." J Biol Chem 287(9);6892-903. PMID: 22223481

Grudniak11: Grudniak AM, Kurek A, Szarlak J, Wolska KI (2011). "Oleanolic and ursolic acids influence affect the expression of the cysteine regulon and the stress response in Escherichia coli." Curr Microbiol 62(4);1331-6. PMID: 21221969

Monroe90: Monroe RS, Ostrowski J, Hryniewicz MM, Kredich NM (1990). "In vitro interactions of CysB protein with the cysK and cysJIH promoter regions of Salmonella typhimurium." J Bacteriol 1990;172(12);6919-29. PMID: 2254265

Ostrowski89a: Ostrowski J, Kredich NM (1989). "Molecular characterization of the cysJIH promoters of Salmonella typhimurium and Escherichia coli: regulation by cysB protein and N-acetyl-L-serine." J Bacteriol 1989;171(1);130-40. PMID: 2701932

Sirko98: Sirko A, Weglenska A, Hulanicka D (1998). "Integration host factor positively regulates cycJIH transcription." Mol Gen Genet 258(1-2);174-7. PMID: 9613586

Zaslaver06: Zaslaver A, Bren A, Ronen M, Itzkovitz S, Kikoin I, Shavit S, Liebermeister W, Surette MG, Alon U (2006). "A comprehensive library of fluorescent transcriptional reporters for Escherichia coli." Nat Methods 3(8);623-8. PMID: 16862137


Report Errors or Provide Feedback
Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
Page generated by SRI International Pathway Tools version 18.5 on Mon Dec 22, 2014, BIOCYC14A.