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dc.contributor.authorEvjenth, Rune H.
dc.contributor.authorBrenner, Annette K.
dc.contributor.authorThompson, Paul R
dc.contributor.authorArnesen, Thomas
dc.contributor.authorFroystein, Nils Age
dc.contributor.authorLillehaug, Johan R.
dc.date2022-08-11T08:11:00.000
dc.date.accessioned2022-08-23T17:28:12Z
dc.date.available2022-08-23T17:28:12Z
dc.date.issued2012-03-23
dc.date.submitted2015-05-22
dc.identifier.citationJ Biol Chem. 2012 Mar 23;287(13):10081-8. doi: 10.1074/jbc.M111.326587. Epub 2012 Feb 6. <a href="http://dx.doi.org/10.1074/jbc.M111.326587">Link to article on publisher's site</a>
dc.identifier.issn0021-9258 (Linking)
dc.identifier.doi10.1074/jbc.M111.326587
dc.identifier.urihttp://hdl.handle.net/20.500.14038/50028
dc.description<p>At the time of publication, Paul Thompson was not yet affiliated with UMass Medical School.</p>
dc.description.abstractN(alpha)-acetylation is a common protein modification catalyzed by different N-terminal acetyltransferases (NATs). Their essential role in the biogenesis and degradation of proteins is becoming increasingly evident. The NAT hNaa50p preferentially modifies peptides starting with methionine followed by a hydrophobic amino acid. hNaa50p also possesses N(epsilon)-autoacetylation activity. So far, no eukaryotic NAT has been mechanistically investigated. In this study, we used NMR spectroscopy, bisubstrate kinetic assays, and product inhibition experiments to demonstrate that hNaa50p utilizes an ordered Bi Bi reaction of the Theorell-Chance type. The NMR results, both the substrate binding study and the dynamic data, further indicate that the binding of acetyl-CoA induces a conformational change that is required for the peptide to bind to the active site. In support of an ordered Bi Bi reaction mechanism, addition of peptide in the absence of acetyl-CoA did not alter the structure of the protein. This model is further strengthened by the NMR results using a catalytically inactive hNaa50p mutant.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=22311970&dopt=Abstract">Link to Article in PubMed</a>
dc.relation.urlhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3323058/
dc.subjectAcetyl Coenzyme A
dc.subjectAcetyltransferases
dc.subjectCatalysis
dc.subjectHumans
dc.subjectHydrophobic and Hydrophilic Interactions
dc.subjectKinetics
dc.subjectMethionine
dc.subject*Models, Chemical
dc.subjectMutation
dc.subjectN-Terminal Acetyltransferase E
dc.subjectNuclear Magnetic Resonance, Biomolecular
dc.subjectPeptides
dc.subjectProtein Conformation
dc.subjectBiochemistry
dc.subjectEnzymes and Coenzymes
dc.subjectMedicinal-Pharmaceutical Chemistry
dc.subjectTherapeutics
dc.titleHuman protein N-terminal acetyltransferase hNaa50p (hNAT5/hSAN) follows ordered sequential catalytic mechanism: combined kinetic and NMR study.
dc.typeJournal Article
dc.source.journaltitleThe Journal of biological chemistry
dc.source.volume287
dc.source.issue13
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/thompson/38
dc.identifier.contextkey7135704
html.description.abstract<p>N(alpha)-acetylation is a common protein modification catalyzed by different N-terminal acetyltransferases (NATs). Their essential role in the biogenesis and degradation of proteins is becoming increasingly evident. The NAT hNaa50p preferentially modifies peptides starting with methionine followed by a hydrophobic amino acid. hNaa50p also possesses N(epsilon)-autoacetylation activity. So far, no eukaryotic NAT has been mechanistically investigated. In this study, we used NMR spectroscopy, bisubstrate kinetic assays, and product inhibition experiments to demonstrate that hNaa50p utilizes an ordered Bi Bi reaction of the Theorell-Chance type. The NMR results, both the substrate binding study and the dynamic data, further indicate that the binding of acetyl-CoA induces a conformational change that is required for the peptide to bind to the active site. In support of an ordered Bi Bi reaction mechanism, addition of peptide in the absence of acetyl-CoA did not alter the structure of the protein. This model is further strengthened by the NMR results using a catalytically inactive hNaa50p mutant.</p>
dc.identifier.submissionpaththompson/38
dc.contributor.departmentDepartment of Biochemistry and Molecular Pharmacology
dc.source.pages10081-8


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