• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of embojLink to Publisher's site
EMBO J. Jun 15, 1999; 18(12): 3204–3213.
PMCID: PMC1171401

Molecular basis of glutathione synthetase deficiency and a rare gene permutation event.

Abstract

Glutathione synthetase (GS) catalyses the production of glutathione from gamma-glutamylcysteine and glycine in an ATP-dependent manner. Malfunctioning of GS results in disorders including metabolic acidosis, 5-oxoprolinuria, neurological dysfunction, haemolytic anaemia and in some cases is probably lethal. Here we report the crystal structure of human GS (hGS) at 2.1 A resolution in complex with ADP, two magnesium ions, a sulfate ion and glutathione. The structure indicates that hGS belongs to the recently identified ATP-grasp superfamily, although it displays no detectable sequence identity with other family members including its bacterial counterpart, Escherichia coli GS. The difficulty in identifying hGS as a member of the family is due in part to a rare gene permutation which has resulted in a circular shift of the conserved secondary structure elements in hGS with respect to the other known ATP-grasp proteins. Nevertheless, it appears likely that the enzyme shares the same general catalytic mechanism as other ligases. The possibility of cyclic permutations provides an insight into the evolution of this family and will probably lead to the identification of new members. Mutations that lead to GS deficiency have been mapped onto the structure, providing a molecular basis for understanding their effects.

Full Text

The Full Text of this article is available as a PDF (442K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Anderson ME. Glutathione: an overview of biosynthesis and modulation. Chem Biol Interact. 1998 Apr 24;111-112:1–14. [PubMed]
  • Artymiuk PJ, Poirrette AR, Rice DW, Willett P. Biotin carboxylase comes into the fold. Nat Struct Biol. 1996 Feb;3(2):128–132. [PubMed]
  • Barton GJ. ALSCRIPT: a tool to format multiple sequence alignments. Protein Eng. 1993 Jan;6(1):37–40. [PubMed]
  • Dahl N, Pigg M, Ristoff E, Gali R, Carlsson B, Mannervik B, Larsson A, Board P. Missense mutations in the human glutathione synthetase gene result in severe metabolic acidosis, 5-oxoprolinuria, hemolytic anemia and neurological dysfunction. Hum Mol Genet. 1997 Jul;6(7):1147–1152. [PubMed]
  • Esser L, Wang CR, Hosaka M, Smagula CS, Südhof TC, Deisenhofer J. Synapsin I is structurally similar to ATP-utilizing enzymes. EMBO J. 1998 Feb 16;17(4):977–984. [PMC free article] [PubMed]
  • Fan C, Moews PC, Walsh CT, Knox JR. Vancomycin resistance: structure of D-alanine:D-alanine ligase at 2.3 A resolution. Science. 1994 Oct 21;266(5184):439–443. [PubMed]
  • Fan C, Moews PC, Shi Y, Walsh CT, Knox JR. A common fold for peptide synthetases cleaving ATP to ADP: glutathione synthetase and D-alanine:d-alanine ligase of Escherichia coli. Proc Natl Acad Sci U S A. 1995 Feb 14;92(4):1172–1176. [PMC free article] [PubMed]
  • Gali RR, Board PG. Sequencing and expression of a cDNA for human glutathione synthetase. Biochem J. 1995 Aug 15;310(Pt 1):353–358. [PMC free article] [PubMed]
  • Gali RR, Board PG. Identification of an essential cysteine residue in human glutathione synthase. Biochem J. 1997 Jan 1;321(Pt 1):207–210. [PMC free article] [PubMed]
  • Galperin MY, Koonin EV. A diverse superfamily of enzymes with ATP-dependent carboxylate-amine/thiol ligase activity. Protein Sci. 1997 Dec;6(12):2639–2643. [PMC free article] [PubMed]
  • Gruer MJ, Artymiuk PJ, Guest JR. The aconitase family: three structural variations on a common theme. Trends Biochem Sci. 1997 Jan;22(1):3–6. [PubMed]
  • Gushima H, Yasuda S, Soeda E, Yokota M, Kondo M, Kimura A. Complete nucleotide sequence of the E. coli glutathione synthetase gsh-II. Nucleic Acids Res. 1984 Dec 21;12(24):9299–9307. [PMC free article] [PubMed]
  • Hara T, Kato H, Katsube Y, Oda J. A pseudo-michaelis quaternary complex in the reverse reaction of a ligase: structure of Escherichia coli B glutathione synthetase complexed with ADP, glutathione, and sulfate at 2.0 A resolution. Biochemistry. 1996 Sep 17;35(37):11967–11974. [PubMed]
  • Jones TA, Zou JY, Cowan SW, Kjeldgaard M. Improved methods for building protein models in electron density maps and the location of errors in these models. Acta Crystallogr A. 1991 Mar 1;47(Pt 2):110–119. [PubMed]
  • Lindqvist Y, Schneider G. Circular permutations of natural protein sequences: structural evidence. Curr Opin Struct Biol. 1997 Jun;7(3):422–427. [PubMed]
  • Lüthy R, Bowie JU, Eisenberg D. Assessment of protein models with three-dimensional profiles. Nature. 1992 Mar 5;356(6364):83–85. [PubMed]
  • Murzin AG. Structural classification of proteins: new superfamilies. Curr Opin Struct Biol. 1996 Jun;6(3):386–394. [PubMed]
  • Murshudov GN, Vagin AA, Dodson EJ. Refinement of macromolecular structures by the maximum-likelihood method. Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240–255. [PubMed]
  • Ristoff E, Larsson A. Patients with genetic defects in the gamma-glutamyl cycle. Chem Biol Interact. 1998 Apr 24;111-112:113–121. [PubMed]
  • Shi ZZ, Habib GM, Rhead WJ, Gahl WA, He X, Sazer S, Lieberman MW. Mutations in the glutathione synthetase gene cause 5-oxoprolinuria. Nat Genet. 1996 Nov;14(3):361–365. [PubMed]
  • Thoden JB, Holden HM, Wesenberg G, Raushel FM, Rayment I. Structure of carbamoyl phosphate synthetase: a journey of 96 A from substrate to product. Biochemistry. 1997 May 27;36(21):6305–6316. [PubMed]
  • Waldrop GL, Rayment I, Holden HM. Three-dimensional structure of the biotin carboxylase subunit of acetyl-CoA carboxylase. Biochemistry. 1994 Aug 30;33(34):10249–10256. [PubMed]
  • Webb GC, Vaska VL, Gali RR, Ford JH, Board PG. The gene encoding human glutathione synthetase (GSS) maps to the long arm of chromosome 20 at band 11.2. Genomics. 1995 Dec 10;30(3):617–619. [PubMed]
  • Whitbread L, Gali RR, Board PG. The structure of the human glutathione synthetase gene. Chem Biol Interact. 1998 Apr 24;111-112:35–40. [PubMed]
  • Wolodko WT, Fraser ME, James MN, Bridger WA. The crystal structure of succinyl-CoA synthetase from Escherichia coli at 2.5-A resolution. J Biol Chem. 1994 Apr 8;269(14):10883–10890. [PubMed]
  • Yamaguchi H, Kato H, Hata Y, Nishioka T, Kimura A, Oda J, Katsube Y. Three-dimensional structure of the glutathione synthetase from Escherichia coli B at 2.0 A resolution. J Mol Biol. 1993 Feb 20;229(4):1083–1100. [PubMed]

Articles from The EMBO Journal are provided here courtesy of The European Molecular Biology Organization

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...