An extended polyanion activation surface in insulin degrading enzyme

Eun Suk Song, Mehmet Ozbil, Tingting Zhang, Michael Sheetz, David Lee, Danny Tran, Sheng Li, Rajeev Prabhakar, Louis B. Hersh, David W. Rodgers

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

Insulin degrading enzyme (IDE) is believed to be the major enzyme that metabolizes insulin and has been implicated in the degradation of a number of other bioactive peptides, including amyloid beta peptide (A?), glucagon, amylin, and atrial natriuretic peptide. IDE is activated toward some substrates by both peptides and polyanions/anions, possibly representing an important control mechanism and a potential therapeutic target. A binding site for the polyanion ATP has previously been defined crystallographically, but mutagenesis studies suggest that other polyanion binding modes likely exist on the same extended surface that forms one wall of the substrate-binding chamber. Here we use a computational approach to define three potential ATP binding sites and mutagenesis and kinetic studies to confirm the relevance of these sites. Mutations were made at four positively charged residues (Arg 429, Arg 431, Arg 847, Lys 898) within the polyanion-binding region, converting them to polar or hydrophobic residues. We find that mutations in all three ATP binding sites strongly decrease the degree of activation by ATP and can lower basal activity and cooperativity. Computational analysis suggests conformational changes that result from polyanion binding as well as from mutating residues involved in polyanion binding. These findings indicate the presence of multiple polyanion binding modes and suggest the anionbinding surface plays an important conformational role in controlling IDE activity.

Original languageEnglish (US)
Article number133114
JournalPloS one
Volume10
Issue number7
DOIs
StatePublished - Jul 17 2015

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

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    Song, E. S., Ozbil, M., Zhang, T., Sheetz, M., Lee, D., Tran, D., Li, S., Prabhakar, R., Hersh, L. B., & Rodgers, D. W. (2015). An extended polyanion activation surface in insulin degrading enzyme. PloS one, 10(7), [133114]. https://doi.org/10.1371/journal.pone.0133114