PXD001437

PXD001437 is an original dataset announced via ProteomeXchange.

Title	N-terminal acetylome analyses reveals the substrate specificity profile of Naa50 (Nat5) and implicate NATs in influencing initiator Methionine processing
Description	Co-translational N-terminal (Nt-) acetylation of nascent polypeptides is catalyzed by N-terminal acetyltransferases (NATs). The very N-terminal amino acid sequence is the major factor determining whether or not a given protein is Nt-acetylated. In humans, six different NATs, denoted NatA-NatF, are identified. In the current study we used N-terminal COFRADIC analysis to define the in vivo substrate specificity of Naa50 (Nat5)/NatE as this has long remained elusive. Three yeast strains were generated; a control strain endogenously expressing yNaa50, a deletion strain depleted of yNaa50 and a strain deleted of yNaa50 but ectopically expressing human Naa50. When comparing the Nt-acetylation status of different N-termini in the control strain with the deletion strain, a reduction in Nt-acetylation for several yeast proteins was observed. To our surprise, these substrates were not of the predicted NatE-type substrates, but rather canonical NatA substrates. Further, ectopic expression of hNaa50 mainly resulted in the Nt-acetylation of a selected class of otherwise Nt-free yeast N-termini besides increasing the degree of Nt-acetylation of several other yeast proteins, and as such (partially) complementing those N-termini displaying reduced Nt-acetylation upon yNAA50-deletion. The preferred substrates of hNaa50 were predominantly Met-starting N-termini including Met-Lys-, Met-Val-, Met-Ala-, Met-Tyr-, Met-Phe-, Met-Leu-, Met-Ser- and Met-Thr, and highly overlapped with the previously identified human Naa60/NatF substrate specificity profile. Identification of several hNaa50 substrates with a small amino acid in the second position also revealed a potential interplay between the NATs and methionine aminopeptidases (MetAPs). The initiator Methionine (iMet) is normally cleaved off by MetAPs when the second amino acid is small, but our in vitro data suggest that in contrast to a free iMet, an Nt-acetylated iMet is not hydrolyzed by MetAPs. Thus, Naa50-mediated Nt-acetylation may potentially act as a mechanism to retain the iMet of proteins with a small amino acid at the second position that normally would be hydrolyzed.
HostingRepository	PRIDE
AnnounceDate	2015-05-29
AnnouncementXML	Submission_2015-05-29_13:13:32.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD001437
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Petra Van Damme
SpeciesList	scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	S-carboxymethyl-L-cysteine: 58.005479; 2xC(13): 3x(2)H labeled N6-acetyl-L-lysine; 2-pyrrolidone-5-carboxylic acid (Gln): -17.026549; N-acetylated residue: 42.010565; L-methionine (S)-sulfoxide: 15.994915
Instrument	LTQ Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2014-10-27 06:45:28	ID requested
⏵ 1	2015-05-29 13:13:33	announced

Van Damme P, Hole K, Gevaert K, Arnesen T, N-terminal acetylome analysis reveals the specificity of Naa50 (Nat5) and suggests a kinetic competition between N-terminal acetyltransferases and methionine aminopeptidases. Proteomics, 15(14):2436-46(2015) [pubmed]

curator keyword: Biological

submitter keyword: N-terminal acetyltransferase / N-terminomics / Nt-acetylome / Naa50 / NatE

Petra Van Damme
contact affiliation	Prof. Petra Van Damme UGent Department of Biochemistry A. Baertsoenkaai 3 B9000 Gent Belgium Tel: +32-92649279
contact email	Petra.vandamme@ugent.be
lab head
Petra Van Damme
contact affiliation	University of Ghent
contact email	petra.vandamme@ugent.be
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD001437
     1. Label: PRIDE project
     2. Name: N-terminal acetylome analyses reveals the substrate specificity profile of Naa50 (Nat5) and implicate NATs in influencing initiator Methionine processing