P0A9D2 (GSTA_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Glutathione S-transferase GstA
EC=2.5.1.18

Alternative name(s):
GST B1-1

Gene names

Name:	gstA
Synonyms:	gst
Ordered Locus Names:	b1635, JW1627

Organism

Escherichia coli (strain K12) [Reference proteome] [HAMAP]

Taxonomic identifier

83333 [NCBI]

Taxonomic lineage

cellular organisms › Bacteria › Proteobacteria › Gammaproteobacteria › Enterobacteriales › Enterobacteriaceae › Escherichia › Escherichia coli

Protein attributes

Sequence length	201 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Catalyzes the conjugation of reduced glutathione (GSH) to a wide number of exogenous and endogenous hydrophobic electrophiles. Shows activity toward 1-chloro-2,4-dinitrobenzene (CDNB) and ethacrynic acid. Also possesses thiol:disulfide oxidoreductase activity, using GSH to reduce bis-(2-hydroxyethyl) disulfide (HEDS). Has a low level of glutathione-dependent peroxidase activity toward cumene hydroperoxide. Is important for defense against oxidative stress, probably via its peroxidase activity. Ref.1 Ref.5 Ref.6 Ref.7
Catalytic activity	RX + glutathione = HX + R-S-glutathione. Ref.1 Ref.5 Ref.7
Subunit structure	Homodimer. Ref.1 Ref.5 Ref.8 Ref.9
Subcellular location	Cytoplasm.
Disruption phenotype	Deletion of the gstA gene decreases the resistance of the bacteria to hydrogen peroxide. Ref.6
Sequence similarities	Belongs to the GST superfamily. Beta family. Contains 1 GST C-terminal domain. Contains 1 GST N-terminal domain.
Biophysicochemical properties	Kinetic parameters: kcat is 9 sec(-1) for the GSH transferase reaction with CDNB as substrate (Ref.7). KM=0.041 mM for glutathione (Ref.1) Ref.1 Ref.5 Ref.7 KM=0.3 mM for glutathione (Ref.7) KM=0.99 mM for 1-chloro-2,4-dinitrobenzene (Ref.1) KM=1.9 mM for 1-chloro-2,4-dinitrobenzene (Ref.7) pH dependence: Optimum pH is 7.5 (Ref.1). Temperature dependence: Optimum temperature is 35 degrees Celsius (Ref.1).

Ontologies

Keywords
Cellular component	Cytoplasm
Molecular function	Transferase
Technical term	3D-structure Complete proteome Direct protein sequencing Reference proteome
Gene Ontology (GO)
Biological_process	xenobiotic catabolic process Traceable author statement PubMed 19016852. Source: EcoCyc
Cellular_component	cytoplasm Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular_function	glutathione transferase activity Inferred from direct assay Ref.1. Source: EcoCyc
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 201

201

Glutathione S-transferase GstA

PRO_0000185970

Regions

Domain

1 – 81

81

GST N-terminal

Domain

87 – 201

115

GST C-terminal

Region

65 – 66

2

Glutathione binding

Region

103 – 106

4

Glutathione binding

Sites

Binding site

10

1

Glutathione

Binding site

35

1

Glutathione

Binding site

52

1

Glutathione; via amide nitrogen and carbonyl oxygen

Binding site

99

1

Glutathione

Experimental info

Mutagenesis

5

1

Y → F: Does not significantly affect the activity. Ref.1

Mutagenesis

10

1

C → A: 8-fold decrease in affinity for GSH, but 5-fold increase in GSH transferase activity. Loss of thiol:disulfide oxidoreductase activity. Ref.7

Mutagenesis

10

1

C → S: 6-fold decrease in affinity for GSH, and decrease in GSH transferase activity. Loss of thiol:disulfide oxidoreductase activity. Ref.7

Mutagenesis

106

1

H → A: Decrease in affinity for GSH while nearly no effect on affinity for CDNB, and decrease in GSH transferase activity. Ref.7

Mutagenesis

106

1

H → F: Decrease in affinity for GSH, while nearly no effect on affinity for CDNB and on GSH transferase activity. Ref.7

Sequence conflict

2

1

K → L AA sequence Ref.5

Sequence conflict

5 – 6

2

YK → IL AA sequence Ref.5

Secondary structure

1

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

201

Helix Strand Turn

Details...

Beta strand

2 – 5

4

Helix

12 – 20

9

Beta strand

26 – 31

6

Turn

32 – 35

4

Helix

43 – 45

3

Beta strand

54 – 56

3

Beta strand

62 – 65

4

Helix

66 – 75

10

Helix

78 – 80

3

Helix

89 – 104

16

Helix

106 – 113

8

Beta strand

115 – 117

3

Helix

119 – 121

3

Helix

122 – 139

18

Turn

140 – 142

3

Beta strand

143 – 145

3

Beta strand

146 – 150

5

Helix

153 – 167

15

Helix

176 – 186

11

Helix

189 – 197

9

Sequences

	Sequence		Length	Mass (Da)
	P0A9D2 [UniParc]. Last modified July 19, 2005. Version 1. Checksum: 6347401123B044E2	FASTA	201	22,868
	10 20 30 40 50 60 MKLFYKPGAC SLASHITLRE SGKDFTLVSV DLMKKRLENG DDYFAVNPKG QVPALLLDDG 70 80 90 100 110 120 TLLTEGVAIM QYLADSVPDR QLLAPVNSIS RYKTIEWLNY IATELHKGFT PLFRPDTPEE 130 140 150 160 170 180 YKPTVRAQLE KKLQYVNEAL KDEHWICGQR FTIADAYLFT VLRWAYAVKL NLEGLEHIAA 190 200 FMQRMAERPE VQDALSAEGL K

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Molecular cloning and site-directed mutagenesis of glutathione S-transferase from Escherichia coli. The conserved tyrosyl residue near the N-terminus is not essential for catalysis." Nishida M., Kong K.-H., Inoue H., Takahashi K. J. Biol. Chem. 269:32536-32541(1994) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-14, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, MUTAGENESIS OF TYR-5. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[2]	"A 570-kb DNA sequence of the Escherichia coli K-12 genome corresponding to the 28.0-40.1 min region on the linkage map." Aiba H., Baba T., Fujita K., Hayashi K., Inada T., Isono K., Itoh T., Kasai H., Kashimoto K., Kimura S., Kitakawa M., Kitagawa M., Makino K., Miki T., Mizobuchi K., Mori H., Mori T., Motomura K. , Nakade S., Nakamura Y., Nashimoto H., Nishio Y., Oshima T., Saito N., Sampei G., Seki Y., Sivasundaram S., Tagami H., Takeda J., Takemoto K., Takeuchi Y., Wada C., Yamamoto Y., Horiuchi T. DNA Res. 3:363-377(1996) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[3]	"The complete genome sequence of Escherichia coli K-12." Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., Shao Y. Science 277:1453-1474(1997) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / MG1655 / ATCC 47076.
[4]	"Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110." Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T. Mol. Syst. Biol. 2:E1-E5(2006) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[5]	"Purification and study of a bacterial glutathione S-transferase." Arca P., Garcia P., Hardisson C., Suarez J.E. FEBS Lett. 263:77-79(1990) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 1-9, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT. Strain: K12.
[6]	"Three distinct-type glutathione S-transferases from Escherichia coli important for defense against oxidative stress." Kanai T., Takahashi K., Inoue H. J. Biochem. 140:703-711(2006) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION IN DEFENSE AGAINST OXIDATIVE STRESS, DISRUPTION PHENOTYPE. Strain: K12.
[7]	"Characterization of the activity and folding of the glutathione transferase from Escherichia coli and the roles of residues Cys(10) and His(106)." Wang X.Y., Zhang Z.R., Perrett S. Biochem. J. 417:55-64(2009) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, CATALYTIC ACTIVITY, KINETIC PARAMETERS, MUTAGENESIS OF CYS-10 AND HIS-106.
[8]	"Three-dimensional structure of Escherichia coli glutathione S-transferase complexed with glutathione sulfonate: catalytic roles of Cys10 and His106." Nishida M., Harada S., Noguchi S., Satow Y., Inoue H., Takahashi K. J. Mol. Biol. 281:135-147(1998) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS) IN COMPLEX WITH GLUTATHIONE SULFONATE, SUBUNIT.
[9]	"Conserved structural elements in glutathione transferase homologues encoded in the genome of Escherichia coli." Rife C.L., Parsons J.F., Xiao G., Gilliland G.L., Armstrong R.N. Proteins 53:777-782(2003) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) IN COMPLEX WITH GLUTATHIONE SULFONATE, SUBUNIT.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

D38497 Genomic DNA. Translation: BAA07509.1.
U00096 Genomic DNA. Translation: AAC74707.1.
AP009048 Genomic DNA. Translation: BAA15396.1.

PIR

A55495.

RefSeq

NP_416152.1. NC_000913.2.
YP_489899.1. NC_007779.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1A0F	X-ray	2.10	A/B	1-201	[»]
1N2A	X-ray	1.90	A/B	1-201	[»]

ProteinModelPortal

P0A9D2.

SMR

P0A9D2. Positions 1-201.

ModBase

Search...

Protein-protein interaction databases

IntAct

P0A9D2. 1 interaction.

STRING

511145.b1635.

2D gel databases

SWISS-2DPAGE

P0A9D2.

Proteomic databases

PaxDb

P0A9D2.

PRIDE

P0A9D2.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

AAC74707; AAC74707; b1635.
BAA15396; BAA15396; BAA15396.

GeneID

12931264.
945758.

KEGG

ecj:Y75_p1612.
eco:b1635.

PATRIC

32118572. VBIEscCol129921_1706.

Organism-specific databases

EchoBASE

EB2497.

EcoGene

EG12613. gstA.

Phylogenomic databases

eggNOG

COG0625.

HOGENOM

HOG000125748.

KO

K00799.

OMA

KFQYVDE.

ProtClustDB

PRK10542.

Enzyme and pathway databases

BioCyc

EcoCyc:GST-MONOMER.
ECOL316407:JW1627-MONOMER.
MetaCyc:GST-MONOMER.

Gene expression databases

Genevestigator

P0A9D2.

Family and domain databases

Gene3D

1.20.1050.10. 1 hit.
3.40.30.10. 1 hit.

InterPro

IPR010987. Glutathione-S-Trfase_C-like.
IPR004045. Glutathione_S-Trfase_N.
IPR017933. Glutathione_S_Trfase/Cl_chnl_C.
IPR004046. GST_C.
IPR012336. Thioredoxin-like_fold.
[Graphical view]

Pfam

PF00043. GST_C. 1 hit.
[Graphical view]

SUPFAM

SSF47616. GST_C_like. 1 hit.
SSF52833. Thiordxn-like_fd. 1 hit.

PROSITE

PS50405. GST_CTER. 1 hit.
PS50404. GST_NTER. 1 hit.
[Graphical view]

ProtoNet

Search...

Other

EvolutionaryTrace

P0A9D2.

Entry information

Entry name

GSTA_ECOLI

Accession

Primary (citable) accession number: P0A9D2
Secondary accession number(s): P39100

Entry history

Integrated into UniProtKB/Swiss-Prot:	July 19, 2005
Last sequence update:	July 19, 2005
Last modified:	May 1, 2013
This is version 68 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program