PXD030189

PXD030189 is an original dataset announced via ProteomeXchange.

Title	A continuous assay set to screen and characterize novel protein N-acetyltransferases unveils rice GNAT2 activity
Description	Protein N-acetyltransferases (NATs) belong to the Gcn5-related N-acetyltransferases (GNATs) superfamily. GNATs catalyze the transfer of acetyl from acetyl-CoA to the reactive amine moiety of a wide range of acceptors. NAT sequences are difficult to distinguish from other members of the GNAT superfamily and there are many uncharacterized GNATs. To facilitate the discovery and characterization of new GNATs, we have developed a new continuous, non-radioactive assay. This assay is virtually independent of the substrate and can be used to get substrate specificity hints. We validated first the assay with the well-characterized Schizosaccharomyces pombe NatA (SpNatA). The SpNatA kinetic parameters were determined with various peptides confirming the robustness of the new assay. We reveal that the longer the peptide substrate the more efficient the enzyme. As a proof of concept of the relevance of the new assay, we characterize a NAA90 member from rice (Oryza sativa), OsGNAT2. We took advantage of an in vivo medium-scale characterization of OsGNAT2 specificity to identify and then validate in vitro several specific peptide substrates. With this assay, we reveal long-range synergic effects of basic residues on OsGNAT2 activity. Overall, this new, high-throughput assay allows better understanding of the substrate specificity and activity of any GNAT.
HostingRepository	PRIDE
AnnounceDate	2022-05-25
AnnouncementXML	Submission_2022-05-25_04:37:57.757.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Jean Baptiste BOYER
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562; scientific name: Oryza sativa (Rice); NCBI TaxID: 4530;
ModificationList	acetylated residue; iodoacetamide derivatized residue
Instrument	LTQ Orbitrap Velos

Revision	Datetime	Status	ChangeLog Entry
0	2021-12-05 07:19:58	ID requested
⏵ 1	2022-05-25 04:37:58	announced

Asensio T, Dian C, Boyer JB, Rivi, è, re F, Meinnel T, Giglione C, A Continuous Assay Set to Screen and Characterize Novel Protein N-Acetyltransferases Unveils Rice General Control Non-repressible 5-Related N-Acetyltransferase2 Activity. Front Plant Sci, 13():832144(2022) [pubmed]

submitter keyword: N-acetyltransferase, acetylation, modifications, GNAT, rice, yeast, NatA

Carmela GIGLIONE
contact affiliation	Protein Maturation, Cell fate and Therapeutics Institute for Integrative, Biology of the Cell (I2BC) CNRS UMR9198, Bâtiment 21, 1 avenue de la Terrasse F-91198 Gif-sur-Yvette cedex, France
contact email	carmela.giglione@i2bc.paris-saclay.fr
lab head
Jean Baptiste BOYER
contact affiliation	Institut de Biologie Intégrative de la Cellule - CNRS
contact email	jean-baptiste.boyer@i2bc.paris-saclay.fr
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2022/04/PXD030189

PRIDE project URI

• PRIDE
  1. PXD030189
     1. Label: PRIDE project
     2. Name: A continuous assay set to screen and characterize novel protein N-acetyltransferases unveils rice GNAT2 activity