Biomolecular NMR Assignments

, Volume 12, Issue 1, pp 163–165 | Cite as

Backbone chemical shift assignments of the glycine cleavage complex H protein of Escherichia coli

Article
  • 83 Downloads

Abstract

Glycine cleavage complex H protein (GcvH) is one of the four components that form the glycine cleavage complex (GCS), essential for the synthesis of C1 (one-carbon units) for cell metabolism, by the oxidative cleavage of glycine. The activity of this complex is induced in the presence of exogenous glycine, and is repressed by purines. GCS, in cooperation with GCA (serine hydroxymethyltransferase) regulates the endogenous levels of glycine and C1 units in the cell. GcvH, the lipoamide containing component of the complex, plays an indispensable role in this reaction, as its prosthetic group shuttles between the active site of the three other components of the GCS complex sequentially. In environments rich in exogenous lipoic acid, GcvH is converted to lipoyl-GcvH by Lipoate protein ligase (LplA), by the salvage pathway. When exogenous lipoic acid is deficient, it is post-translationally modified to lipoyl-GcvH by the consecutive action of two enzymes, (a) Lipoate protein ligase B (LipB) and (b) Lipoyl synthase (LipA). Although, the crystal structure has been determined for Escherichia coli GcvH, no information exists for its interaction with LipB or LipA. Therefore, we plan to study its interactions with the aforementioned enzymes. As a first step, we have carried out the complete backbone chemical shift assignments of the E. coli glycine cleavage complex H protein in its apo-form, as well as its C8- intermediate.

Keywords

Glycine cleavage complex H protein Glycine cleavage complex Nuclear magnetic resonance Backbone resonance assignments Lipoic acid 

Notes

Acknowledgements

We thank the Department of Biotechnology (DBT), Govt. of India for financial and infrastructure support. Fellowship to Usha Yadav from University Grants commission (UGC), India is also thankfully acknowledged.

References

  1. Cao X, Cronan JE (2015) The Streptomyces coelicolor lipoate-protein ligase is a circularly permuted version of the Escherichia coli enzyme composed of discrete interacting domains. J Biol Chem 290(11):7280–7290CrossRefGoogle Scholar
  2. Cronan JE (2014) Biotin and lipoic acid: synthesis, attachment, and regulation. EcoSal Plus 6(1):1–39Google Scholar
  3. Cronan JE (2016) Assembly of lipoic acid on its cognate enzymes: an extraordinary and essential biosynthetic pathway. Microbiol Mol Biol Rev 80(2):429–450CrossRefGoogle Scholar
  4. Cronan JE, Zhao X et al (2005) Function, attachment and synthesis of lipoic acid in Escherichia coli. Adv Microb Physiol 50:103–146CrossRefGoogle Scholar
  5. Delaglio F, Grzesiek S et al (1995) NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6(3):277–293CrossRefGoogle Scholar
  6. Jordan SW, Cronan JE (2003) The Escherichia coli lipB gene encodes lipoyl (octanoyl)-acyl carrier protein:protein transferase. J Bacteriol 185(5):1582–1589CrossRefGoogle Scholar
  7. Keller RLJ (2004) Optimizing the process of nuclear magnetic resonance spectrum analysis and computer aided resonance assignment. Dissertation. Swiss Federal Institute of Technology, ZurichGoogle Scholar
  8. Macherel D, Bourguignon J et al (1996) Expression, lipoylation and structure determination of recombinant pea H-protein in Escherichia coli. Eur J Biochem 236:27–33CrossRefGoogle Scholar
  9. Morris TW, Reed KE et al (1994) Identification of the gene encoding lipoate-protein ligase A of Escherichia coli. Molecular cloning and characterization of the lplA gene and gene product. J Biol Chem 269(23):16091–16100Google Scholar
  10. Morris TW, Reed KE et al (1995) Lipoic acid metabolism in Escherichia coli: the lplA and lipB genes define redundant pathways for ligation of lipoyl groups to apoprotein. J Bacteriol 177(1):1–10CrossRefGoogle Scholar
  11. Nesbitt NM, Baleanu-Gogonea C et al (2005) Expression, purification, and physical characterization of Escherichia coli lipoyl(octanoyl) transferase. Protein Exp Purif 39(2):269–282CrossRefGoogle Scholar
  12. Okamura-Ikeda K, Ohmura Y et al (1993) Cloning and nucleotide sequence of the gcv operon encoding the Escherichia coli glycine-cleavage system. Eur J Biochem 216(2):539–548CrossRefGoogle Scholar
  13. Wilson RL, Steiert PS et al (1993) Positive regulation of the Escherichia coli glycine cleavage enzyme system. J Bacteriol 175(3):902–904CrossRefGoogle Scholar
  14. Zhao X, Miller JR et al (2005) The reaction of LipB, the octanoyl-[acyl carrier protein]:protein N-octanoyltransferase of lipoic acid synthesis, proceeds through an acyl-enzyme intermediate. Biochemistry 44(50):16737–16746CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.National Institute of ImmunologyAruna Asaf Ali MargNew DelhiIndia

Personalised recommendations