DB code: M00051

RLCP classification 3.113.90030.1133 : Transfer
3.1143.90000.1132 : Transfer
CATH domain 3.30.1490.80 : Dna Ligase; domain 1
3.30.470.20 : D-amino Acid Aminotransferase; Chain A, domain 1 Catalytic domain
1.10.1080.10 : Glutathione Synthetase; Chain A, domain 3 Catalytic domain
3.40.50.1760 : Rossmann fold
3.30.1490.50 : Dna Ligase; domain 1 Catalytic domain
E.C. 6.3.2.3
CSA 2hgs
M-CSA 2hgs
MACiE

CATH domain Related DB codes (homologues)
3.30.470.20 : D-amino Acid Aminotransferase; Chain A, domain 1 T00082 D00298 M00035 M00037 T00107 T00108

Uniprot Enzyme Name
UniprotKB Protein name Synonyms RefSeq Pfam
P48637 Glutathione synthetase
EC 6.3.2.3
Glutathione synthase
GSH synthetase
GSH-S
NP_000169.1 (Protein)
NM_000178.2 (DNA/RNA sequence)
PF03917 (GSH_synth_ATP)
PF03199 (GSH_synthase)
[Graphical View]
Q08220 Glutathione synthetase
EC 6.3.2.3
Glutathione synthase
GSH synthetase
GSH-S
NP_014593.1 (Protein)
NM_001183303.1 (DNA/RNA sequence)
PF03917 (GSH_synth_ATP)
PF03199 (GSH_synthase)
[Graphical View]

KEGG enzyme name
glutathione synthase
glutathione synthetase
GSH synthetase

UniprotKB: Accession Number Entry name Activity Subunit Subcellular location Cofactor
P48637 GSHB_HUMAN ATP + gamma-L-glutamyl-L-cysteine + glycine = ADP + phosphate + glutathione. Homodimer (By similarity).
Q08220 GSHB_YEAST ATP + gamma-L-glutamyl-L-cysteine + glycine = ADP + phosphate + glutathione.

KEGG Pathways
Map code Pathways E.C.
MAP00251 Glutamate metabolism
MAP00480 Glutathione metabolism

Compound table
Cofactors Substrates Products Intermediates
KEGG-id C00305 C00669 C00002 C00037 C00008 C00009 C00051
E.C.
Compound Magnesium gamma-L-Glutamyl-L-cysteine ATP Glycine ADP Orthophosphate Glutathione
Type divalent metal (Ca2+, Mg2+) amino acids,amide group,sulfhydryl group amine group,nucleotide amino acids amine group,nucleotide phosphate group/phosphate ion amino acids,carboxyl group,peptide/protein,sulfhydryl group
ChEBI 18420
 18420
 		17515
 17515
 		15422
 15422
 		15428
 57305
 15428
 57305
 		16761
 16761
 		26078
 26078
 		16856
 16856
PubChem 888
 888
 		123938
 123938
 		5957
 5957
 		5257127
 750
 5257127
 750
 		6022
 6022
 		1004
 22486802
 1004
 22486802
 		124886
 25246407
 124886
 25246407
2hgsA01 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1m0tA01 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1m0tB01 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1m0wA01 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1m0wB01 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
2hgsA02 Bound:_MG Unbound Unbound Unbound Unbound Analogue:SO4_505 Bound:GSH
1m0tA02 Unbound Unbound Unbound Unbound Unbound Analogue:SO4_504 Unbound
1m0tB02 Unbound Unbound Unbound Unbound Unbound Analogue:SO4_503 Unbound
1m0wA02 Bound:_MG Analogue:3GC Analogue:ANP Unbound Unbound Unbound Unbound
1m0wB02 Bound:_MG Analogue:3GC Analogue:ANP Unbound Unbound Unbound Unbound
2hgsA03 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1m0tA03 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1m0tB03 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1m0wA03 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1m0wB03 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
2hgsA04 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1m0tA04 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1m0tB04 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1m0wA04 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1m0wB04 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
2hgsA05 Bound:_MG Unbound Unbound Unbound Bound:ADP Unbound Unbound
1m0tA05 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1m0tB05 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1m0wA05 Bound:_MG Unbound Unbound Unbound Unbound Unbound Unbound
1m0wB05 Bound:_MG Unbound Unbound Unbound Unbound Unbound Unbound

Reference for Active-site residues
resource references E.C.
literature [6] & [12]

Active-site residues
PDB Catalytic residues Cofactor-binding residues Modified residues Main-chain involved in catalysis Comment
2hgsA01
1m0tA01
1m0tB01
1m0wA01
1m0wB01
2hgsA02 ARG 125;ARG 450 GLU 144(Mg1 & Mg2 binding);ASN 146(Mg2 binding) SER 151
1m0tA02 ARG 128;ARG 467 GLU 146(Mg1 & Mg2 binding);ASN 148(Mg2 binding) SER 153
1m0tB02 ARG 1128;ARG 1467 GLU 1146(Mg1 & Mg2 binding);ASN 1148(Mg2 binding) SER 1153
1m0wA02 ARG 128;ARG 467 GLU 146(Mg1 & Mg2 binding);ASN 148(Mg2 binding) SER 153
1m0wB02 ARG 1128;ARG 1467 GLU 1146(Mg1 & Mg2 binding);ASN 1148(Mg2 binding) SER 1153
2hgsA03 LYS 305
1m0tA03 LYS 324
1m0tB03 LYS 1324
1m0wA03 LYS 324
1m0wB03 LYS 1324
2hgsA04
1m0tA04
1m0tB04
1m0wA04
1m0wB04
2hgsA05 LYS 364 GLU 368(Mg2 binding) GLY 369
1m0tA05 LYS 382 invisible 385-388
1m0tB05 LYS 1382 invisible 1385-1389
1m0wA05 LYS 382 GLU 386(Mg2 binding) GLY 387
1m0wB05 LYS 1382 GLU 1386(Mg2 binding) invisible 1387-1388

References for Catalytic Mechanism
References Sections No. of steps in catalysis
[6]
p.3208-3209
[12]

References
[1]
Resource
Comments
Medline ID
PubMed ID 447639
Journal J Biol Chem
Year 1979
Volume 254
Pages 5184-90
Authors Oppenheimer L, Wellner VP, Griffith OW, Meister A
Title Glutathione synthetase. Purification from rat kidney and mapping of the substrate binding sites.
Related PDB
Related UniProtKB
[2]
Resource
Comments
Medline ID
PubMed ID 7181863
Journal Biochem J
Year 1982
Volume 207
Pages 65-72
Authors York MJ, Kuchel PW, Chapman BE, Jones AJ
Title Incorporation of labelled glycine into reduced glutathione of intact human erythrocytes by enzyme-catalysed exchange. A nuclear-magnetic-resonance study.
Related PDB
Related UniProtKB
[3]
Resource
Comments
Medline ID
PubMed ID 6148935
Journal Biomed Biochim Acta
Year 1984
Volume 43
Pages 719-26
Authors Kuchel PW, Chapman BE, Endre ZH, King GF, Thorburn DR, York MJ
Title Monitoring metabolic reactions in erythrocytes using NMR spectroscopy.
Related PDB
Related UniProtKB
[4]
Resource
Comments
Medline ID
PubMed ID 7567920
Journal Protein Eng
Year 1995
Volume 8
Pages 353-62
Authors Mizuguchi K, Go N
Title Comparison of spatial arrangements of secondary structural elements in proteins.
Related PDB
Related UniProtKB
[5]
Resource
Comments Homologous enzyme
Medline ID
PubMed ID 9551557
Journal Structure
Year 1998
Volume 6
Pages 363-76
Authors Levdikov VM, Barynin VV, Grebenko AI, Melik-Adamyan WR, Lamzin VS, Wilson KS
Title The structure of SAICAR synthase: an enzyme in the de novo pathway of purine nucleotide biosynthesis.
Related PDB
Related UniProtKB
[6]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 10369661
Journal EMBO J
Year 1999
Volume 18
Pages 3204-13
Authors Polekhina G, Board PG, Gali RR, Rossjohn J, Parker MW
Title Molecular basis of glutathione synthetase deficiency and a rare gene permutation event.
Related PDB 2hgs
Related UniProtKB
[7]
Resource
Comments
Medline ID
PubMed ID 10438618
Journal J Mol Biol
Year 1999
Volume 291
Pages 239-47
Authors Grishin NV
Title Phosphatidylinositol phosphate kinase: a link between protein kinase and glutathione synthase folds.
Related PDB
Related UniProtKB
[8]
Resource
Comments
Medline ID
PubMed ID 10861239
Journal Biochem J
Year 2000
Volume 349
Pages 275-9
Authors Njalsson R, Carlsson K, Olin B, Carlsson B, Whitbread L, Polekhina G, Parker MW, Norgren S, Mannervik B, Board PG, Larsson A
Title Kinetic properties of missense mutations in patients with glutathione synthetase deficiency.
Related PDB
Related UniProtKB
[9]
Resource
Comments
Medline ID
PubMed ID 11964186
Journal Biochem J
Year 2002
Volume 363
Pages 833-8
Authors Meierjohann S, Walter RD, Muller S
Title Glutathione synthetase from Plasmodium falciparum.
Related PDB
Related UniProtKB
[10]
Resource
Comments
Medline ID
PubMed ID 12467574
Journal Structure (Camb)
Year 2002
Volume 10
Pages 1669-76
Authors Gogos A, Shapiro L
Title Large conformational changes in the catalytic cycle of glutathione synthase.
Related PDB 1m0t 1m0w
Related UniProtKB
[11]
Resource
Comments
Medline ID
PubMed ID 12734194
Journal J Biol Chem
Year 2003
Volume 278
Pages 40152-61
Authors Phlippen N, Hoffmann K, Fischer R, Wolf K, Zimmermann M
Title The glutathione synthetase of Schizosaccharomyces pombe is synthesized as a homodimer but retains full activity when present as a heterotetramer.
Related PDB
Related UniProtKB
[12]
Resource
Comments
Medline ID
PubMed ID 14990577
Journal J Biol Chem
Year 2004
Volume 279
Pages 22412-21
Authors Dinescu A, Cundari TR, Bhansali VS, Luo JL, Anderson ME
Title Function of conserved residues of human glutathione synthetase: implications for the ATP-grasp enzymes.
Related PDB
Related UniProtKB

Comments
According to the literature [6], this enzyme catalyzes two successive transfer reactions. Firstly, it transfers the gamma-phosphate group of ATP to the C-terminal carboxylate of the second substrate, gamma-glutamylcysteine, to form an acylphosphate intermediate. Secondly, it transfers the acyl group from the intermediate to the amine group of the third substrate, glycine, to form a tetrahedral carbon inermediate, which dissociates into the product GSH, releasing inorganic phosphate and ADP.
The first reaction (phosphoryl transfer) proceeds as follows (see [6]):
(1) The acceptor group, the C-terminal carboxylate oxygen of gamma-glutamylcysteine (the first substrate), makes a nucleophilic attack on the transferred group, gamma-phosphate of ATP (the second substrate), leading to the formation of the pentacovalent phosphate transition state.
(2) The mainchain amide of Gly369, and the sidechains of Arg125 and Arg450 stabilize the transferred group, gamma-phosphate, together with two magnesium ions bound to Glu144, Asn146 and Glu368, during the transition state. Meanwhile, Lys305 and Lys364 stabilize the negative charge of the leaving group, alpha- and beta-phosphate groups of ATP (the second substrate), together with the two magnesium ions.
(3) The leaving group, ADP, dissociates, forming an acylphosphate intermediate.
The second reaction (acyl transfer) proceeds as follows (see [6] & [12]):
(1') The acceptor group, the amine of glycine, makes a nucleophilic attack on the transferred group, the carbonyl carbon of the acylphosphate intermediate, forming the tetrahedral intermediate.
(2') The mainchain amide of Ser151, and the sidechain of Arg125 stabilize the charge on the tetrahedral intermediate. Meanwhile, Arg125 and Arg450 stabilize the negative charge of the leaving gamma-phosphate, together with the two magnesium ions.
(3') Finally, the tetrahedral carbon intermediate dissociates to form the product GSH, releasing the inorganic phosphate.

Created Updated
2004-08-01 2009-03-16