Reviewed, UniProtKB/Swiss-Prot P04425 (GSHB_ECOLI)

Names and origin

Protein names

Recommended name:
    Glutathione synthetase
    EC=6.3.2.3
Alternative name(s):
    Glutathione synthase
    GSH synthetase
      Short name=GSH-S
      Short name=GSHase

Gene names

Name:	gshB
Synonyms:	gsh-II
Ordered Locus Names:	b2947, JW2914

Organism

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

Taxonomic identifier

83333 [NCBI]

Taxonomic lineage

Bacteria › Proteobacteria › Gammaproteobacteria › Enterobacteriales › Enterobacteriaceae › Escherichia

Protein attributes

Sequence length	316 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is not processed.
Protein existence	Evidence at protein level.

General annotation (Comments)

Catalytic activity	ATP + gamma-L-glutamyl-L-cysteine + glycine = ADP + phosphate + glutathione. HAMAP MF_00162
Cofactor	Binds 1 magnesium or manganese ion per subunit By similarity.
Enzyme regulation	Inhibited by 7,8-dihydrofolate, methotrexate and trimethoprim. HAMAP MF_00162
Pathway	Sulfur metabolism; glutathione biosynthesis; glutathione from L-cysteine and L-glutamate: step 2/2. HAMAP MF_00162
Subunit structure	Homotetramer. HAMAP MF_00162
Sequence similarities	Belongs to the prokaryotic GSH synthase family. Contains 1 ATP-grasp domain.

Ontologies

Keywords
Biological process	Glutathione biosynthesis
Ligand	ATP-binding Magnesium Manganese Metal-binding Nucleotide-binding
Molecular function	Ligase
Technical term	3D-structure Complete proteome Direct protein sequencing
Gene Ontology (GO)
Biological process	glutathione biosynthetic process Inferred from electronic annotation. Source: HAMAP
Cellular component	cytosol Inferred from direct assay. Source: UniProtKB
Molecular function	ATP binding Inferred from electronic annotation. Source: HAMAP glutathione synthase activity Inferred from electronic annotation. Source: HAMAP magnesium ion binding Inferred from electronic annotation. Source: UniProtKB-KW manganese ion binding Inferred from electronic annotation. Source: UniProtKB-KW protein binding Inferred from physical interaction. Source: IntAct
Complete GO annotation...

Binary interactions

With	Entry	#Exp.	IntAct	Notes
map	P0AE18	1	EBI-909107,EBI-553355
nadE	P18843	1	EBI-909107,EBI-548960

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 316

316

Glutathione synthetase HAMAP MF_00162

PRO_0000197465

Regions

Domain

125 – 310

186

ATP-grasp

Nucleotide binding

151 – 207

57

ATP By similarity

Sites

Metal binding

281

1

Magnesium or manganese By similarity

Metal binding

283

1

Magnesium or manganese By similarity

Experimental info

Mutagenesis

122

1

C → A: No loss of activity. Ref.8

Mutagenesis

195

1

C → A: No loss of activity. Ref.8

Mutagenesis

222

1

C → A: No loss of activity. Ref.8

Mutagenesis

227

1

P → V: Loss of activity. Ref.8 Ref.6

Mutagenesis

233

1

R → A or K: Loss of activity. Ref.8 Ref.7

Mutagenesis

240

1

G → V: Loss of activity. Ref.8 Ref.6

Mutagenesis

241

1

R → A or K: No loss of activity. Ref.8 Ref.7

Mutagenesis

289

1

C → A: No loss of activity. Ref.8

Secondary structure

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316

Helix Strand Turn

Details...

Beta strand

3 – 9

7

Helix

11 – 13

3

Turn

16 – 18

3

Helix

20 – 30

11

Beta strand

34 – 38

5

Helix

40 – 42

3

Beta strand

43 – 46

4

Beta strand

49 – 59

11

Beta strand

67 – 76

10

Helix

77 – 79

3

Beta strand

81 – 86

6

Helix

93 – 107

15

Beta strand

111 – 114

4

Helix

116 – 121

6

Turn

124 – 126

3

Helix

127 – 130

4

Helix

132 – 134

3

Beta strand

138 – 142

5

Helix

144 – 154

11

Beta strand

155 – 161

7

Beta strand

171 – 173

3

Helix

180 – 187

8

Turn

188 – 192

5

Beta strand

195 – 199

5

Helix

202 – 206

5

Beta strand

208 – 214

7

Beta strand

220 – 246

27

Helix

249 – 264

16

Beta strand

269 – 275

7

Beta strand

278 – 283

6

Helix

290 – 296

7

Helix

301 – 314

14

Sequences

	Sequence		Length	Mass (Da)
	P04425-1 [UniParc]. Last modified August 13, 1987. Version 1. Checksum: F2E67AB29DE113DD	FASTA	316	35,561
	10 20 30 40 50 60 MIKLGIVMDP IANINIKKDS SFAMLLEAQR RGYELHYMEM GDLYLINGEA RAHTRTLNVK 70 80 90 100 110 120 QNYEEWFSFV GEQDLPLADL DVILMRKDPP FDTEFIYATY ILERAEEKGT LIVNKPQSLR 130 140 150 160 170 180 DCNEKLFTAW FSDLTPETLV TRNKAQLKAF WEKHSDIILK PLDGMGGASI FRVKEGDPNL 190 200 210 220 230 240 GVIAETLTEH GTRYCMAQNY LPAIKDGDKR VLVVDGEPVP YCLARIPQGG ETRGNLAAGG 250 260 270 280 290 300 RGEPRPLTES DWKIARQIGP TLKEKGLIFV GLDIIGDRLT EINVTSPTCI REIEAEFPVS 310 ITGMLMDAIE ARLQQQ

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Complete nucleotide sequence of the E. coli glutathione synthetase gsh-II." Gushima H., Yasuda S., Soeda E., Yokota M., Kondo M., Kimura A. Nucleic Acids Res. 12:9299-9307(1984) [PubMed: 6393055] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: B.
[2]	"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: 9278503] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / MG1655 / ATCC 47076.
[3]	"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: 16738553] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[4]	"Three-dimensional structure of the glutathione synthetase from Escherichia coli B at 2.0-A resolution." Yamaguchi H., Kato H., Hata Y., Nishioka T., Kimura A., Oda J., Katsube Y. J. Mol. Biol. 229:1083-1100(1993) [PubMed: 8445637] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS). Strain: B.
[5]	"Crystal structure of glutathione synthetase at optimal pH: domain architecture and structural similarity with other proteins." Matsuda K., Mizuguchi K., Nishioka T., Kato H., Go N., Oda J. Protein Eng. 9:1083-1092(1996) [PubMed: 9010922] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS). Strain: B.
[6]	"Flexibility impaired by mutations revealed the multifunctional roles of the loop in glutathione synthetase." Tanaka T., Yamaguchi H., Kato H., Nishioka T., Katsube Y., Oda J. Biochemistry 32:12398-12404(1993) [PubMed: 8241129] [Abstract] Cited for: MUTAGENESIS OF PRO-227 AND GLY-240.
[7]	"Mutational and proteolytic studies on a flexible loop in glutathione synthetase from Escherichia coli B: the loop and arginine 233 are critical for the catalytic reaction." Tanaka T., Kato H., Nishioka T., Oda J. Biochemistry 31:2259-2265(1992) [PubMed: 1540581] [Abstract] Cited for: PROTEIN SEQUENCE OF 234-242, MUTAGENESIS OF ARG-233 AND ARG-241. Strain: B.
[8]	"Role of cysteine residues in glutathione synthetase from Escherichia coli B. Chemical modification and oligonucleotide site-directed mutagenesis." Kato H., Tanaka T., Nishioka T., Kimura A., Oda J. J. Biol. Chem. 263:11646-11651(1988) [PubMed: 3042775] [Abstract] Cited for: MUTAGENESIS OF CYSTEINE RESIDUES. Strain: B.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBLGenBankDDBJ

X01666 Genomic DNA. Translation: CAA25826.1.
U28377 Genomic DNA. Translation: AAA69114.1.
U00096 Genomic DNA. Translation: AAC75984.1.
AP009048 Genomic DNA. Translation: BAE77010.1.

PIR

SYECGS. A01194.

RefSeq

AP_003504.1.
NP_417422.1.

3D structure databases

PDBPDBe

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1GLV	X-ray	2.70	A	1-316	[»]
1GSA	X-ray	2.00	A	1-316	[»]
1GSH	X-ray	2.00	A	1-316	[»]
2GLT	X-ray	2.20	A	1-316	[»]

ModBase

Search...

Protein-protein interaction databases

DIP

DIP:9840N.

IntAct

P04425. 4 interactions.

Genome annotation databases

GeneID

947445.

GenomeReviews

Gene locus JW2914 in contig AP009048_GR.
Gene locus b2947 in contig U00096_GR.

KEGG

ecj:JW2914.
eco:b2947.

Organism-specific databases

EchoBASE

EB0414.

EcoGene

EG10419. gshB.

CMR

Search...

Phylogenomic databases

HOGENOM

P04425.

OMA

P04425. VRNAPEK.

Enzyme and pathway databases

BioCyc

EcoCyc:GLUTATHIONE-SYN-MON.
MetaCyc:GLUTATHIONE-SYN-MON.

Family and domain databases

HAMAP

MF_00162.
[Tree]

InterPro

IPR011761. ATP-grasp.
IPR013816. ATP_grasp_subdomain_2.
IPR006284. Glut_synth_pro.
IPR004218. GSHS_ATP_bd.
IPR004215. GSHS_N.
IPR013817. Pre-ATP_grasp.
[Graphical view]

Gene3D

G3DSA:3.30.470.20. ATP_grasp_subdomain_2. 1 hit.
G3DSA:3.40.50.20. Pre-ATP_grasp. 1 hit.

Pfam

PF02955. GSH-S_ATP. 1 hit.
PF02951. GSH-S_N. 1 hit.
[Graphical view]

TIGRFAMs

TIGR01380. glut_syn. 1 hit.

PROSITE

PS50975. ATP_GRASP. 1 hit.
[Graphical view]

ProtoNet

Search...

Entry information

Entry name

GSHB_ECOLI

Accession

Primary (citable) accession number: P04425
Secondary accession number(s): Q2M9P6

Entry history

Integrated into UniProtKB/Swiss-Prot:	August 13, 1987
Last sequence update:	August 13, 1987
Last modified:	June 16, 2009
This is version 94 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

HAMAP (High-quality Automated and Manual Annotation of microbial Proteomes)