MetaCyc Enzyme: glyoxalase III, Hsp31 molecular chaperone

Gene: hchA Accession Numbers: G7055 (MetaCyc), b1967, ECK1963

Synonyms: yzzC, yedU, Hsp31

Species: Escherichia coli K-12 substr. MG1655

Subunit composition of glyoxalase III, Hsp31 molecular chaperone = [HchA]2
         glyoxalase III, Hsp31 molecular chaperone = HchA

Glyoxalase III converts methylglyoxal to D-lactate in a single glutathione (GSH)-independent step. The other known route for this conversion is the two-step GSH-dependent pathway catalyzed by glyoxalase I and II, methylglyoxal degradation I. Glyoxalase III was partially characterized by [Misra95], although the gene encoding the enzyme was not identified at that time. It was recently shown that glyoxalase III is identical to Hsp31 in E. coli [Subedi11].

Active site residues have been predicted [Wei07]. Mutations in predicted residues of the catalytic triad, E77A, C185A and H186A, strongly reduce catalytic activity of the enzyme [Subedi11].

Hsp31 was first identified as a chaperone that interacts with partially and fully denatured proteins [Sastry02, Malki03]. At elevated temperatures, Hsp31 homodimers bind to substrates undergoing denaturation, and upon temperature decrease the chaperone dissociates from the renatured substrates [Sastry02]. Hsp31 appears to have weak proteolytic activity using BSA as a substrate [Lee03]; others have found no endo- or exopeptidase activity [Malki03, Zhao03]. Later, Hsp31 was shown to have an aminopeptidase activity with short peptide substrates that is abolished in a C185A mutant [Malki05].

Hsp31 may bind ATP, but does not hydrolyze it [Sastry02, Malki03]. ATP induces conformational changes within Hsp31 and is inhibitory with respect to chaperone activity at high temperatures [Sastry02].

Each Hsp31 monomer comprises two domains separated by a flexible linker region [Quigley03, Quigley04]. Crystal structures are presented at 1.6 Å [Quigley03], 2.2 Å [Zhao03], 2.7 Å [Quigley04] and 2.8 Å [Lee03] resolution. Analysis of a variety of mutants identified the linker-loop region of Hsp31 as a gate which may control access of nonnative proteins to the high-affinity substrate binding site and which is thermally activated [Sastry04]. Extensions of Hsp31 with histidine tags at either its N- or C-terminus abolish chaperone activity [Sastry09].

An hchA mutant has a growth defect at 48°C and is strongly affected in its ability to recover from transient exposure to 50°C [Mujacic04]. In stationary phase, an hchA mutant is more sensitive to methylglyoxal [Subedi11] and acid stress [Mujacic07] than wild type.

Unlike glyoxalases I and II, glyoxalase III activity increases at the transition to stationary phase; the increase is dependent on RpoS [Benov04]. Glyoxalase III activity is not induced by growth in the presence of methylglyoxal [OkadoMatsumoto00]. Expression of hchA is induced by heat shock [Richmond99a]. hchA is a member of the σS stress regulon [Mujacic06], and Hsp31 expression is also induced by osmotic stress [Weber06]. The stability of the hchA transcript is higher at increased temperature [Rasouly07].

Citations: [Kim02]

Locations: cytosol

Map Position: [2,033,859 -> 2,034,710]

Molecular Weight of Polypeptide: 31.19 kD (from nucleotide sequence), 31 kD (experimental) [Sastry02 ]

Molecular Weight of Multimer: 62.0 kD (experimental) [Sastry02]

Unification Links: ASAP:ABE-0006529 , DIP:DIP-11851N , EchoBASE:EB1705 , EcoGene:EG11755 , EcoliWiki:b1967 , ModBase:P31658 , OU-Microarray:b1967 , PortEco:hchA , PR:PRO_000022864 , Pride:P31658 , Protein Model Portal:P31658 , RefSeq:NP_416476 , RegulonDB:G7055 , SMR:P31658 , String:511145.b1967 , UniProt:P31658

Relationship Links: InterPro:IN-FAMILY:IPR002818 , InterPro:IN-FAMILY:IPR017283 , InterPro:IN-FAMILY:IPR029062 , PDB:Structure:1IZY , PDB:Structure:1IZZ , PDB:Structure:1N57 , PDB:Structure:1ONS , PDB:Structure:1PV2 , Pfam:IN-FAMILY:PF01965

Gene-Reaction Schematic: ?

Gene-Reaction Schematic

GO Terms:

Biological Process: GO:0010447 - response to acidic pH Inferred from experiment [Mujacic07]
GO:0019243 - methylglyoxal catabolic process to D-lactate Inferred from experiment [Subedi11]
GO:0019249 - lactate biosynthetic process Inferred from experiment [Subedi11]
GO:0051595 - response to methylglyoxal Inferred from experiment [Subedi11]
GO:0042026 - protein refolding Inferred by computational analysis [GOA06, GOA et al., 2001]
Molecular Function: GO:0005515 - protein binding Inferred from experiment [Rajagopala14, Arifuzzaman06]
GO:0019172 - glyoxalase III activity Inferred from experiment [Subedi11]
GO:0008270 - zinc ion binding Inferred by computational analysis [GOA06]
GO:0016787 - hydrolase activity Inferred by computational analysis [UniProt-GOA, 2011]
GO:0016829 - lyase activity Inferred by computational analysis [UniProt-GOA, 2011, GOA et al., 2001]
GO:0046872 - metal ion binding Inferred by computational analysis [UniProt-GOA, 2011]
GO:0051082 - unfolded protein binding Inferred by computational analysis [GOA06]
Cellular Component: GO:0005829 - cytosol Inferred from experiment Inferred by computational analysis [Diaz-Mejia et al., 2009, Ishihama et al., 2008, Lopez-Campistrous et al., 2005, Lasserre et al., 2006]
GO:0005737 - cytoplasm Inferred by computational analysis [UniProt-GOA, 2011a, UniProt-GOA, 2011, GOA06]

MultiFun Terms: cell processes protection detoxification
information transfer protein related chaperoning, repair (refolding)

Imported from EcoCyc 27-Jan-2015 by Paley S , SRI International

Enzymatic reaction of: glyoxalase

EC Number:

methylglyoxal + H2O <=> (R)-lactate + H+

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the direction of enzyme catalysis.

The reaction is irreversible in the direction shown.

Alternative Substrates for methylglyoxal: phenylglyoxal [Misra95 ]

In Pathways: superpathway of methylglyoxal degradation , methylglyoxal degradation II

Imported from EcoCyc 27-Jan-2015 by Paley S , SRI International

Inhibitors (Unknown Mechanism): Zn2+ [Subedi11] , Cu2+ [Subedi11] , p-hydroxymercuribenzoate [Misra95, Comment 1] , 5,5'-dithio-bis-2-nitrobenzoate [Misra95, Comment 1] , N-ethylmaleimide [Misra95, Comment 1]

Kinetic Parameters:

Km (μM)
kcat (sec-1)
kcat/Km (sec-1 μM-1)

T(opt): 35-40 °C [Subedi11]

pH(opt): 6-8 [Misra95]

Sequence Features

Feature Class Location Citations Comment
Cleavage-of-Initial-Methionine 1
[Yoshida93, UniProt, 2011]
UniProt: Removed.
Chain 2 -> 283
[UniProt, 2009]
UniProt: Chaperone protein hchA;
Mutagenesis-Variant 77
[Subedi11, UniProt12b]
UniProt: Almost completely abolishes glyoxalase activity.
Metal-Binding-Site 86
[UniProt, 2010]
UniProt: Zinc;
Metal-Binding-Site 91
[UniProt, 2010]
UniProt: Zinc;
Metal-Binding-Site 123
[UniProt, 2010]
UniProt: Zinc;
Mutagenesis-Variant 185
[Lee03, Subedi11, UniProt12b]
UniProt: Almost completely abolishes glyoxalase and aminopeptidase activities.
Active-Site 185
[UniProt, 2010a]
UniProt: Non-Experimental Qualifier: probably;
Mutagenesis-Variant 186
[Subedi11, UniProt12b]
UniProt: Shows approximately 17% remaining glyoxalase activity compared with that of the wild-type.
Active-Site 186
[UniProt, 2010a]
UniProt: Non-Experimental Qualifier: probably;
Active-Site 214
[UniProt, 2010a]
UniProt: Non-Experimental Qualifier: probably;

Peter D. Karp on Thu Jan 16, 2003:
Predicted gene function revised as a result of E. coli genome reannotation by Serres et al. [Serres01 ].
Markus Krummenacker on Tue Oct 14, 1997:
Gene object created from Blattner lab Genbank (v. M52) entry.


Arifuzzaman06: Arifuzzaman M, Maeda M, Itoh A, Nishikata K, Takita C, Saito R, Ara T, Nakahigashi K, Huang HC, Hirai A, Tsuzuki K, Nakamura S, Altaf-Ul-Amin M, Oshima T, Baba T, Yamamoto N, Kawamura T, Ioka-Nakamichi T, Kitagawa M, Tomita M, Kanaya S, Wada C, Mori H (2006). "Large-scale identification of protein-protein interaction of Escherichia coli K-12." Genome Res 16(5);686-91. PMID: 16606699

Benov04: Benov L, Sequeira F, Beema AF (2004). "Role of rpoS in the regulation of glyoxalase III in Escherichia coli." Acta Biochim Pol 51(3);857-60. PMID: 15448747

Diaz-Mejia et al., 2009: 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

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

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

Ishihama et al., 2008: Ishihama Y, Schmidt T, Rappsilber J, Mann M, Hartl FU, Kerner MJ, Frishman D (2008). "Protein abundance profiling of the Escherichia coli cytosol." BMC Genomics 9;102. PMID: 18304323

Kim02: Kim OG, Kim IK, Kim GH, Ko J, Park C, Suh PG, Kang SO, Lee HS, Cha SS (2002). "Crystallization and preliminary X-ray crystallographic analysis of a yedU gene product from Escherichia coli." Acta Crystallogr D Biol Crystallogr 58(Pt 7);1217-9. PMID: 12077448

Lasserre et al., 2006: Lasserre JP, Beyne E, Pyndiah S, Lapaillerie D, Claverol S, Bonneu M (2006). "A complexomic study of Escherichia coli using two-dimensional blue native/SDS polyacrylamide gel electrophoresis." Electrophoresis 27(16);3306-21. PMID: 16858726

Lee03: Lee SJ, Kim SJ, Kim IK, Ko J, Jeong CS, Kim GH, Park C, Kang SO, Suh PG, Lee HS, Cha SS (2003). "Crystal structures of human DJ-1 and Escherichia coli Hsp31, which share an evolutionarily conserved domain." J Biol Chem 278(45);44552-9. PMID: 12939276

Lopez-Campistrous et al., 2005: Lopez-Campistrous A, Semchuk P, Burke L, Palmer-Stone T, Brokx SJ, Broderick G, Bottorff D, Bolch S, Weiner JH, Ellison MJ (2005). "Localization, annotation, and comparison of the Escherichia coli K-12 proteome under two states of growth." Mol Cell Proteomics 4(8);1205-9. PMID: 15911532

Malki03: Malki A, Kern R, Abdallah J, Richarme G (2003). "Characterization of the Escherichia coli YedU protein as a molecular chaperone." Biochem Biophys Res Commun 301(2);430-6. PMID: 12565879

Malki05: Malki A, Caldas T, Abdallah J, Kern R, Eckey V, Kim SJ, Cha SS, Mori H, Richarme G (2005). "Peptidase activity of the Escherichia coli Hsp31 chaperone." J Biol Chem 280(15);14420-6. PMID: 15550391

Misra95: Misra K, Banerjee AB, Ray S, Ray M (1995). "Glyoxalase III from Escherichia coli: a single novel enzyme for the conversion of methylglyoxal into D-lactate without reduced glutathione." Biochem J 1995;305 ( Pt 3);999-1003. PMID: 7848303

Mujacic04: Mujacic M, Bader MW, Baneyx F (2004). "Escherichia coli Hsp31 functions as a holding chaperone that cooperates with the DnaK-DnaJ-GrpE system in the management of protein misfolding under severe stress conditions." Mol Microbiol 51(3);849-59. PMID: 14731284

Mujacic06: Mujacic M, Baneyx F (2006). "Regulation of Escherichia coli hchA, a stress-inducible gene encoding molecular chaperone Hsp31." Mol Microbiol 60(6);1576-89. PMID: 16796689

Mujacic07: Mujacic M, Baneyx F (2007). "Chaperone Hsp31 contributes to acid resistance in stationary-phase Escherichia coli." Appl Environ Microbiol 73(3);1014-8. PMID: 17158627

OkadoMatsumoto00: Okado-Matsumoto A, Fridovich I (2000). "The role of alpha,beta -dicarbonyl compounds in the toxicity of short chain sugars." J Biol Chem 275(45);34853-7. PMID: 10931845

Quigley03: Quigley PM, Korotkov K, Baneyx F, Hol WG (2003). "The 1.6-A crystal structure of the class of chaperones represented by Escherichia coli Hsp31 reveals a putative catalytic triad." Proc Natl Acad Sci U S A 100(6);3137-42. PMID: 12621151

Quigley04: Quigley PM, Korotkov K, Baneyx F, Hol WG (2004). "A new native EcHsp31 structure suggests a key role of structural flexibility for chaperone function." Protein Sci 13(1);269-77. PMID: 14691241

Rajagopala14: Rajagopala SV, Sikorski P, Kumar A, Mosca R, Vlasblom J, Arnold R, Franca-Koh J, Pakala SB, Phanse S, Ceol A, Hauser R, Siszler G, Wuchty S, Emili A, Babu M, Aloy P, Pieper R, Uetz P (2014). "The binary protein-protein interaction landscape of Escherichia coli." Nat Biotechnol 32(3);285-90. PMID: 24561554

Rasouly07: Rasouly A, Shenhar Y, Ron EZ (2007). "Thermoregulation of Escherichia coli hchA transcript stability." J Bacteriol 189(15);5779-81. PMID: 17526696

Richmond99a: Richmond CS, Glasner JD, Mau R, Jin H, Blattner FR (1999). "Genome-wide expression profiling in Escherichia coli K-12." Nucleic Acids Res 27(19);3821-35. PMID: 10481021

Sastry02: Sastry MS, Korotkov K, Brodsky Y, Baneyx F (2002). "Hsp31, the Escherichia coli yedU gene product, is a molecular chaperone whose activity is inhibited by ATP at high temperatures." J Biol Chem 277(48);46026-34. PMID: 12235139

Sastry04: Sastry MS, Quigley PM, Hol WG, Baneyx F (2004). "The linker-loop region of Escherichia coli chaperone Hsp31 functions as a gate that modulates high-affinity substrate binding at elevated temperatures." Proc Natl Acad Sci U S A 101(23);8587-92. PMID: 15173574

Sastry09: Sastry MS, Zhou W, Baneyx F (2009). "Integrity of N- and C-termini is important for E. coli Hsp31 chaperone activity." Protein Sci 18(7);1439-47. PMID: 19517531

Serres01: Serres MH, Gopal S, Nahum LA, Liang P, Gaasterland T, Riley M (2001). "A functional update of the Escherichia coli K-12 genome." Genome Biol 2(9);RESEARCH0035. PMID: 11574054

Subedi11: Subedi KP, Choi D, Kim I, Min B, Park C (2011). "Hsp31 of Escherichia coli K-12 is glyoxalase III." Mol Microbiol 81(4);926-36. PMID: 21696459

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

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

UniProt, 2010a: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 00:00:00." Database.

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

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

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

UniProt12b: UniProt Consortium (2012). "UniProt version 2012-09 released on 2012-09-12 00:00:00." Database.

Weber06: Weber A, Kogl SA, Jung K (2006). "Time-dependent proteome alterations under osmotic stress during aerobic and anaerobic growth in Escherichia coli." J Bacteriol 188(20);7165-75. PMID: 17015655

Wei07: Wei Y, Ringe D, Wilson MA, Ondrechen MJ (2007). "Identification of functional subclasses in the DJ-1 superfamily proteins." PLoS Comput Biol 3(1);e10. PMID: 17257049

Yoshida93: Yoshida T, Ueguchi C, Yamada H, Mizuno T (1993). "Function of the Escherichia coli nucleoid protein, H-NS: molecular analysis of a subset of proteins whose expression is enhanced in a hns deletion mutant." Mol Gen Genet 237(1-2);113-22. PMID: 8455549

Zhao03: Zhao Y, Liu D, Kaluarachchi WD, Bellamy HD, White MA, Fox RO (2003). "The crystal structure of Escherichia coli heat shock protein YedU reveals three potential catalytic active sites." Protein Sci 12(10);2303-11. PMID: 14500888

Report Errors or Provide Feedback
Please cite the following article in publications resulting from the use of MetaCyc: Caspi et al, Nucleic Acids Research 42:D459-D471 2014
Page generated by SRI International Pathway Tools version 19.0 on Mon May 25, 2015, BIOCYC13B.