title: Purified recombinant hARD1 does not catalyse acetylation of Lys532 of HIF-1alpha fragments in vitro.
abstract: In humans, many responses to hypoxia including angiogenesis and erythropoiesis are mediated by the alpha/beta-heterodimeric transcription factor hypoxia inducible factor (HIF). The stability and/or activity of human HIF-1alpha are modulated by post-translational modifications including prolyl and asparaginyl hydroxylation, phosphorylation, and reportedly by acetylation of the side-chain of Lys532 by ARD1 (arrest defective protein 1 homologue), an acetyltransferase. Using purified recombinant human ARD1 (hARD1) we did not observe ARD1-mediated N-acetylation of Lys532 using fragments of HIF-1alpha. However, recombinant hARD1 from Escherichia coli was produced with partial N-terminal acetylation and was observed to undergo slow self-mediated N-terminal acetylation. The observations are consistent with the other data indicating that hARD1, at least alone, does not acetylate HIF-1alpha, and with reports on the N-terminal acetyltransferase activity of a recently reported heterodimeric complex comprising hARD1 and N-acetyltransferase protein.
authors.affiliation: The Department of Chemistry and The Oxford Centre for Molecular Sciences, Chemistry Research Laboratory, University of Oxford, Mansfield Road, Oxford OX1 3TA, United Kingdom.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Murray-Rust
authors.forename: Neil J
authors.initials: NJ
authors.lastname: Oldham
authors.forename: Kirsty S
authors.initials: KS
authors.lastname: Hewitson
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Schofield
mesh_terms: D000107:Acetylation; D000123:Acetyltransferases / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D000324:Adrenocorticotropic Hormone / Q000378:metabolism; D002384:Catalysis; D002853:Chromatography, Liquid; D006639:Histidine / Q000378:metabolism; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism*; D008239:Lysine / Q000378:metabolism*; D063209:N-Terminal Acetyltransferase A; D063213:N-Terminal Acetyltransferase E; D009842:Oligopeptides / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: C471213:His-His-His-His-His-His; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D006639:Histidine; D000324:Adrenocorticotropic Hormone; D000123:Acetyltransferases; D063209:N-Terminal Acetyltransferase A; C504824:NAA10 protein, human; D063213:N-Terminal Acetyltransferase E; D008239:Lysine