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ATP-citrate lyase multimerization is required for coenzyme-A substrate binding and catalysis.

Gleb A Bazilevsky1,2, Hayley C Affronti2,3, Xuepeng Wei2,4

  • 1From the Graduate Group in Cell and Molecular Biology.

The Journal of Biological Chemistry
|March 17, 2019
PubMed
Summary

The C-terminal domain of ATP-citrate lyase (ACLY) is essential for enzyme tetramerization and CoA binding, crucial for acetyl-CoA production. This finding reveals a new regulatory mechanism for ACLY, impacting cellular metabolism and disease.

Keywords:
ATP-citrate lyaseacetyl coenzyme A (acetyl-CoA)citrate synthasecoenzyme A (CoA)enzyme mechanismmetabolismprotein assembly

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cellular Metabolism

Background:

  • ATP-citrate lyase (ACLY) is a key enzyme producing nucleocytosolic acetyl-CoA, vital for eukaryotic carbon metabolism.
  • Dysregulation of ACLY is implicated in cancers, cardiovascular diseases, and metabolic disorders.
  • The precise molecular mechanisms governing ACLY activity and function remain incompletely understood.

Purpose of the Study:

  • To investigate the role of the uncharacterized C-terminal citrate synthase homology domain of ACLY in acetyl-CoA formation.
  • To elucidate the structural and functional significance of the ACLY C terminus in enzyme activity.

Main Methods:

  • Purification of recombinant ACLY.
  • Biochemical and biophysical assays including analytical ultracentrifugation, dynamic light scattering, and thermal stability assays.
  • Assessment of CoA binding and enzyme activity in the context of the full-length enzyme and its C-terminal domain.

Main Results:

  • The ACLY C terminus is critical for maintaining enzyme homotetramerization, a necessary quaternary structure.
  • The C-terminal domain, within the full-length enzyme, is essential for optimal CoA binding.
  • ACLY tetramerization mediated by the C terminus facilitates efficient acetyl-CoA production.

Conclusions:

  • The C-terminal citrate synthase homology domain plays a novel and essential role in ACLY function and catalysis.
  • ACLY forms a homotetramer via its C terminus to enable CoA binding and acetyl-CoA synthesis.
  • This discovery offers insights into ACLY regulation with potential therapeutic implications for targeting acetyl-CoA-dependent processes.