Trypanosomatid Deoxyhypusine Synthase Activity Is Dependent on Shared Active-Site Complementation between Pseudoenzyme Paralogs

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Trypanosoma brucei is a neglected tropical disease endemic to Africa. The polyamine spermidine is essential for post-translational hypusine modification of eukaryotic initiation factor 5A (eIF5A), which is catalyzed by deoxyhypusine synthase (TbDHS). In trypanosomatids, deoxyhypusine synthase (DHS) activity is dependent on heterotetramer formation between two paralogs, DHSc and DHSp, both with minimal activity on their own due to missing catalytic residues. We determined the X-ray structure of TbDHS showing a single functional shared active site is formed at the DHSc/DHSp heterodimer interface, with deficiencies in one subunit complemented by the other. Each heterodimer contains two NAD+ binding sites, one housed in the functional catalytic site and the second bound in a remnant dead site that lacks key catalytic residues. Functional analysis of these sites by site-directed mutagenesis identified long-range contributions to the catalytic site from the dead site. Differences between trypanosomatid and human DHS that could be exploited for drug discovery were identified. Afanador et al. determined the X-ray structure of an essential enzyme for survival of the eukaryotic pathogen Trypanosoma brucei. This enzyme is composed of two inactive paralogs, together forming an active heterotetramer via active-site complementation. This structure differs from the human homolog, highlighting the potential for species-selective inhibition.

Original languageEnglish (US)
Pages (from-to)1499-1512.e5
JournalStructure
Volume26
Issue number11
DOIs
StatePublished - Nov 6 2018

Fingerprint

Catalytic Domain
Trypanosoma brucei brucei
X-Rays
Neglected Diseases
Eukaryotic Initiation Factors
Spermidine
Polyamines
Enzymes
Drug Discovery
Post Translational Protein Processing
Site-Directed Mutagenesis
NAD
Binding Sites
Survival
deoxyhypusine synthase

Keywords

  • deoxyhypusine synthase
  • eIF5A
  • hypusine
  • polyamines
  • pseudoenzyme
  • Trypanosoma brucei
  • trypanosomatids

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

Trypanosomatid Deoxyhypusine Synthase Activity Is Dependent on Shared Active-Site Complementation between Pseudoenzyme Paralogs. / Afanador, Gustavo A.; Tomchick, Diana R; Phillips, Margaret A.

In: Structure, Vol. 26, No. 11, 06.11.2018, p. 1499-1512.e5.

Research output: Contribution to journalArticle

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