Transition-state analysis of 2-O acetyl-ADP-ribose hydrolysis by human macrodomain 1

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Macrodomains, including the human macrodomain 1 (MacroD1), are erasers of the post-translational modification of monoadenosinediphospho-ribosylation and hydrolytically deacetylate the sirtuin product O-acetyl-ADP-ribose (OAADPr). OAADPr has been reported to play a role in cell signaling based on oocyte microinjection studies, and macrodomains affect an array of cell processes including transcription and response to DNA damage. Here, we investigate human MacroD1 by transition-state (TS) analysis based on kinetic isotope effects (KIEs) from isotopically labeled OAADPr substrates. Competitive radiolabeled-isotope effects and mass spectrometry were used to obtain KIE data to yield intrinsic KIE values. Intrinsic KIEs were matched to a quantum chemical structure of the TS that includes the active site residues Asp<sup>184</sup> and Asn<sup>174</sup> and a structural water molecule. Transition-state analysis supports a concerted mechanism with an early TS involving simultaneous nucleophilic water attack and leaving group bond cleavage where the breaking C-O ester bond = 1.60 A˚ and the C-O bond to the attacking water nucleophile = 2.30 A˚. The MacroD1 TS provides mechanistic understanding of the OAADPr esterase chemistry. (Chemical Presented).

Original languageEnglish (US)
Pages (from-to)2255-2262
Number of pages8
JournalACS Chemical Biology
Volume9
Issue number10
DOIs
StatePublished - Oct 17 2014

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O-Acetyl-ADP-Ribose
Isotopes
Hydrolysis
Kinetics
Water
Cell signaling
Nucleophiles
Microinjections
Esterases
Transcription
Post Translational Protein Processing
DNA Damage
Oocytes
Mass spectrometry
Mass Spectrometry
Catalytic Domain
Esters
2-O-acetyl-ADP-ribose
Molecules
DNA

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

Cite this

Transition-state analysis of 2-O acetyl-ADP-ribose hydrolysis by human macrodomain 1. / Hirsch, Brett M.; Burgos, Emmanuel S.; Schramm, Vern L.

In: ACS Chemical Biology, Vol. 9, No. 10, 17.10.2014, p. 2255-2262.

Research output: Contribution to journalArticle

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