The chemical basis of serine palmitoyltransferase inhibition by myriocin

John M. Wadsworth, David J. Clarke, Stephen A. McMahon, Jonathan P. Lowther, Ashley E. Beattie, Pat R. R. Langridge-Smith, Howard B. Broughton, Teresa M. Dunn, James H. Naismith, Dominic J. Campopiano*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

66 Citations (Scopus)

Abstract

Sphingolipids (SLs) are essential components of cellular membranes formed from the condensation of L-serine and a long-chain acyl thioester. This first step is catalyzed by the pyridoxal-5'-phosphate (PLP)-dependent, enzyme serine palmitoyltransferase (SPT) which is a promising therapeutic target. The fungal natural product myriocin is a potent inhibitor of SPT and is widely used to block SL biosynthesis despite a lack of a detailed understanding of its molecular-mechanism. By combining spectroscopy, mass spectrometry, X-ray crystallography, and kinetics, we have characterized the molecular details of SPT inhibition by myriocin. Myriocin initially forms an external aldimine with PLP at the active site, and a structure of the resulting co-complex explains its nanomolar affinity for the enzyme. This co-complex then catalytically degrades via an unexpected 'retro-aldol-like' cleavage mechanism to a C18 aldehyde which in turn acts as a suicide inhibitor of SPT by covalent modification of the essential catalytic lysine. This surprising dual mechanism of inhibition rationalizes the extraordinary potency and longevity of myriocin inhibition.

Original languageEnglish
Pages (from-to)14276-14285
Number of pages10
JournalJournal of the American Chemical Society
Volume135
Issue number38
DOIs
Publication statusPublished - 25 Sept 2013

Keywords

  • PYRIDOXAL-PHOSPHATE ENZYMES
  • SPHINGOLIPID BIOSYNTHESIS
  • REACTION-MECHANISM
  • CRYSTAL-STRUCTURE
  • DRUG DISCOVERY
  • KEY ENZYME
  • AMINO-ACID
  • PROTEINS
  • CRYSTALLOGRAPHY
  • INACTIVATION

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