The Mechanism of N-Terminal Acetylation of Protein

Driessen, H.P.C.; Jong, W.W. de; Tesser, G. I.; Bloemendal, H.

doi:10.3109/10409238509086784

Cited by 223 publications

(137 citation statements)

References 136 publications

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“…The fact that the PEXEL motifs of both exported proteins are cleaved and N-acetylated prior to ERto-Golgi transport establishes that both PEXEL processing events occur in the ER. Since there is precedence that N-terminal cleavage and acetylation of eukaryotic proteins are cotranslational events [29,30], PEXEL processing could be co-translational as exported proteins are translocated across the parasite ER membrane.…”

Section: Pexel Processing Occurs In the Parasite Ermentioning

confidence: 99%

N-terminal processing of proteins exported by malaria parasites

Chang

Falick

Carlton

et al. 2008

Molecular and Biochemical Parasitology

171

177

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Malaria parasites utilize a short N-terminal amino acid motif termed the Plasmodium export element (PEXEL) to export an array of proteins to the host erythrocyte during blood stage infection. Using immunoaffinity chromatography and mass spectrometry, insight into this signalmediated trafficking mechanism was gained by discovering that the PEXEL motif is cleaved and N-acetylated. PfHRPII and PfEMP2 are two soluble proteins exported by Plasmodium falciparum that were demonstrated to undergo PEXEL cleavage and N-acetylation, thus indicating that this Nterminal processing may be general to many exported soluble proteins. It was established that PEXEL processing occurs upstream of the brefeldin A-sensitive trafficking step in the P. falciparum secretory pathway, therefore cleavage and N-acetylation of the PEXEL motif occurs in the endoplasmic reticulum (ER) of the parasite. Furthermore, it was shown that the recognition of the processed N-terminus of exported proteins within the parasitophorous vacuole may be crucial for protein transport to the host erythrocyte. It appears that the PEXEL may be defined as a novel ER peptidase cleavage site and a classical N-acetyltransferase substrate sequence.

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Section: Pexel Processing Occurs In the Parasite Ermentioning

confidence: 99%

N-terminal processing of proteins exported by malaria parasites

Chang

Falick

Carlton

et al. 2008

Molecular and Biochemical Parasitology

171

177

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“…In eukaryotes, N-terminal acetylation, a stable modification of processed (ϪMet) and unprocessed (ϩMet) protein N termini by addition of an acetyl moiety, is a widely observed modification that occurs cotranslationally, subsequent to the opportunity for methionine processing (45). Unlike methionine processing, a simple relationship between N-terminal sequence and modification state has not been found (14,44,45).…”

Section: N-terminal Modificationsmentioning

confidence: 99%

Direct mass spectrometric analysis of intact proteins of the yeast large ribosomal subunit using capillary LC/FTICR

Lee

Berger

Martinović

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

182

133

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Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry coupled with capillary reverse-phase liquid chromatography was used to characterize intact proteins from the large subunit of the yeast ribosome. High mass measurement accuracy, achieved by ''mass locking'' with an internal standard from a dual electrospray ionization source, allowed identification of ribosomal proteins. Analyses of the intact proteins revealed information on cotranslational and posttranslational modifications of the ribosomal proteins that included loss of the initiating methionine, acetylation, methylation, and proteolytic maturation. High-resolution separations permitted differentiation of protein isoforms having high structural similarity as well as proteins from their modified forms, facilitating unequivocal assignments. The study identified 42 of the 43 core large ribosomal subunit proteins and 58 (of 64 possible) core large subunit protein isoforms having unique masses in a single analysis. These results demonstrate the basis for the high-throughput analyses of complex mixtures of intact proteins, which we believe will be an important complement to other approaches for defining protein modifications and their changes resulting from physiological processes or environmental perturbations.M ass spectrometry has evolved into a powerful tool for analyzing biomolecules because of the development of matrix-assisted laser desorption ionization (1, 2) and electrospray ionization (ESI) (3) and advances in both mass analyzers and data processing capabilities. With these ionization methods, which are amenable to megadalton-size molecules, the broadly useful detection and characterization of biopolymers by mass spectrometric methods are feasible. For example, MS has become a preferred analytical tool for proteome analyses, in which dynamic populations of proteins are identified and quantified. Proteins are now routinely identified by mass spectrometric and tandem mass spectrometric analysis of proteolytic digests of individual protein spots on two-dimensional (2D) PAGE (4). However, the intact protein level analyses of complex protein mixtures, while potentially providing complementary and direct information for protein identification, have been a much greater experimental challenge and only rarely attempted (5, 6).Many pivotal cellular processes are not carried out by individual proteins, but rather by large protein-protein complexes and protein-nucleic acid complexes. The analyses of such complexes have been greatly expanded by improvements in both biological separations and MS. One of the more complicated protein complexes yet studied by MS is the cytosolic ribosome complex. Efforts to define the protein composition (7, 8), modifications (9), and subunit interactions (10) of ribosomes generally have mirrored the development of biomolecular analysis techniques and experimental approaches for the study of noncovalent protein complexes.Ribosomes are the canonical example of ribonucleoprotein complexes in both prokaryot...

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“…Studies using 2D-PAGE, HPLC separations, and mass spectrometry revealed that Ϸ50% of all cytosolic yeast proteins are N-acetylated (1). For mammalian proteins, early studies and database searches revealed that 80% to 90% are N-acetylated (9)(10)(11). However, more recent studies, including small-scale experiments with several mammalian proteins, indicated that this number may be closer to 30% (12).…”

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confidence: 99%

Proteomics analyses reveal the evolutionary conservation and divergence of N-terminal acetyltransferases from yeast and humans

Arnesen

Damme

Polevoda

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

496

716

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N ␣ -terminal acetylation is one of the most common protein modifications in eukaryotes. The COmbined FRActional DIagonal Chromatography (COFRADIC) proteomics technology that can be specifically used to isolate N-terminal peptides was used to determine the N-terminal acetylation status of 742 human and 379 yeast protein N termini, representing the largest eukaryotic dataset of N-terminal acetylation. The major N-terminal acetyltransferase (NAT), NatA, acts on subclasses of proteins with Ser-, Ala-, Thr-, Gly-, Cys-and Val-N termini. NatA is composed of subunits encoded by yARD1 and yNAT1 in yeast and hARD1 and hNAT1 in humans. A yeast ard1-⌬ nat1-⌬ strain was phenotypically complemented by hARD1 hNAT1, suggesting that yNatA and hNatA are similar. However, heterologous combinations, hARD1 yNAT1 and yARD1 hNAT1, were not functional in yeast, suggesting significant structural subunit differences between the species. Proteomics of a yeast ard1-⌬ nat1-⌬ strain expressing hNatA demonstrated that hNatA acts on nearly the same set of yeast proteins as yNatA, further revealing that NatA from humans and yeast have identical or nearly identical specificities. Nevertheless, all NatA substrates in yeast were only partially N-acetylated, whereas the corresponding NatA substrates in HeLa cells were mainly completely N-acetylated. Overall, we observed a higher proportion of N-terminally acetylated proteins in humans (84%) as compared with yeast (57%). N-acetylation occurred on approximately one-half of the human proteins with Met-Lys-termini, but did not occur on yeast proteins with such termini. Thus, although we revealed different N-acetylation patterns in yeast and humans, the major NAT, NatA, acetylates the same substrates in both species.Ard1 ͉ COFRADIC ͉ N-terminal acetylation ͉ Nat1 ͉ NatA P rotein N ␣ -terminal acetylation (here referred to as Nacetylation) is one of the most common covalent modifications of eukaryotic proteins, in which an acetyl group is transferred from acetyl-CoA to the ␣-amino group of protein N-terminal residues. N-acetylation occurs cotranslationally on nascent polypeptide chains and almost all N-acetylations in Saccharomyces cerevisiae are catalyzed by 1 of 3 major Nterminal acetyltransferase (NAT) complexes, NatA, NatB or NatC, consisting of catalytic subunits Ard1p, Nat3p, and Mak3p, respectively, and 1 or more auxiliary subunits (1). Yeast NatA, the major and best studied NAT, is composed of the catalytic subunit Ard1p in complex with Nat1p (2). Nat1p is responsible for anchoring Ard1p to the ribosome, thus facilitating cotranslational N-acetylation (3). Both subunits are required for optimal acetyltransferase activity and yeast strains lacking either one of the subunits display the same phenotypes, indicating that both genes are also functionally linked (4). The yeast NatA, NatB and NatC complexes differ in their substrate specificities. NatA substrates represent by far the largest group and contain proteins with Ser-, Ala-, Gly-, Val-, Cys-or Thr-N termini, whereas NatB and NatC act on diff...

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The Mechanism of N-Terminal Acetylation of Protein

Cited by 223 publications

References 136 publications

N-terminal processing of proteins exported by malaria parasites

N-terminal processing of proteins exported by malaria parasites

Direct mass spectrometric analysis of intact proteins of the yeast large ribosomal subunit using capillary LC/FTICR

Proteomics analyses reveal the evolutionary conservation and divergence of N-terminal acetyltransferases from yeast and humans

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