N-terminal protein modifications: Bringing back into play the ribosome

Giglione, Carmela; Fieulaine, Sonia; Meinnel, Thierry

doi:10.1016/j.biochi.2014.11.008

Cited by 159 publications

(168 citation statements)

References 173 publications

Supporting

Mentioning

165

Contrasting

Order By: Relevance

“…NatE complex that is composed of Naa10, Naa15, and Naa50 is reported to be associated with the ribosome (16). From earlier reports, it is clear that Naa10 acetylates the amino termini of all peptides with small and uncharged amino acids (5,8). In the present study, we have shown that human Naa50 acetylates the methionine followed by any residue except for proline.…”

Section: Broad Substrate Specificity Of Nate May Account For Majoritysupporting

confidence: 58%

“…Based on these studies, it was established that different NAT complexes have preferences for different amino termini. For example, NatA, which is a complex of Naa10 and Naa15, has been implicated in acetylating peptides that have small and uncharged amino acids at the amino terminus that are formed after the removal of initiator methionine by MetAP (8). Similarly, Naa50 acetylates the amino-terminal methionine that is followed by leucine, isoleucine, phenylalanine, and tryptophan (8,13).…”

Section: Resultsmentioning

confidence: 99%

“…For example, NatA, which is a complex of Naa10 and Naa15, has been implicated in acetylating peptides that have small and uncharged amino acids at the amino terminus that are formed after the removal of initiator methionine by MetAP (8). Similarly, Naa50 acetylates the amino-terminal methionine that is followed by leucine, isoleucine, phenylalanine, and tryptophan (8,13). Because our x-ray crystal structure suggests Met-Met in the first two positions, we designed a series of tetrapeptides to explore the substrate specificity of Naa50.…”

Section: Resultsmentioning

confidence: 99%

“…NatA acetylates the peptides with serine, alanine, threonine, cysteine, and valine on the amino termini that are formed after MetAP action at the ribosome exit tunnel (5,9), whereas NatB acts on methionine followed by acidic residues in the second position (10,11). NatC, NatE, and NatF act on methionine that is followed by hydrophobic amino acids like leucine, phenylalanine, isoleucine, and tryptophan (8,10,12,13). Additionally, NatF also acetylates methionine that is followed specifically by a lysine (14).…”

mentioning

confidence: 99%

See 3 more Smart Citations

Human Naa50 Protein Displays Broad Substrate Specificity for Amino-terminal Acetylation

Reddi¹,

Saddanapu²,

Chinthapalli

et al. 2016

Journal of Biological Chemistry

View full text Add to dashboard Cite

Amino-terminal acetylation is a critical co-translational modification of the newly synthesized proteins in a eukaryotic cell carried out by six amino-terminal acetyltransferases (NATs). All NATs contain at least one catalytic subunit, and some contain one or two additional auxiliary subunits. For example, NatE is a complex of Naa10, Naa50, and Naa15 (auxiliary). In the present study, the crystal structure of human Naa50 suggested the presence of CoA and acetylated tetrapeptide (AcMMXX) that have co-purified with the protein. Biochemical and thermal stability studies on the tetrapeptide library with variations in the first and second positions confirm our results from the crystal structure that a peptide with Met-Met in the first two positions is the best substrate for this enzyme. In addition, Naa50 acetylated all MXAA peptides except for MPAA. Transcriptome analysis of 10 genes that make up six NATs in humans from eight different cell lines suggests that components of NatE are transcribed in all cell lines, whereas others are variable. Because Naa10 is reported to acetylate all amino termini that are devoid of methionine and Naa50 acetylates all other peptides that are followed by methionine, we believe that NatE complex can be a major contributor for amino-terminal acetylation at the ribosome exit tunnel.Co-translational protein modifications are important biochemical events. Prominent among them are amino-terminal modifications such as methionine cleavage, acetylation, and myristoylation (1-3). Initiator methionine in eukaryotic proteins is removed co-translationally by the enzyme methionine aminopeptidase (MetAP) 3 when the second amino acid is small and uncharged (1). About 70% of the matured proteins do not retain their amino-terminal methionine (4). Similarly, about 90% of all newly synthesized cytosolic proteins undergo aminoterminal acetylation (5). Protein amino-terminal acetylation is carried out by six amino-terminal acetyltransferases (NATs) named sequentially NatA to NatF, classified based on substrate preference (6 -8). NatA acetylates the peptides with serine, alanine, threonine, cysteine, and valine on the amino termini that are formed after MetAP action at the ribosome exit tunnel (5, 9), whereas NatB acts on methionine followed by acidic residues in the second position (10, 11). NatC, NatE, and NatF act on methionine that is followed by hydrophobic amino acids like leucine, phenylalanine, isoleucine, and tryptophan (8, 10, 12, 13). Additionally, NatF also acetylates methionine that is followed specifically by a lysine (14). NatD acetylates the amino termini of Ser-Gly of histones H2A and H4 (15). Each of the NATs (NatA-NatF) is a complex of one or two of the enzymes labeled serially as Naa10, Naa20, Naa30, Naa40, Naa50, and Naa60 attached to one or two of the auxiliary proteins named Naa15, Naa25, Naa35, and Naa38 (7, 9). Some NATs are known to bind to the ribosome at the exit tunnel through their auxiliary subunits (11, 16). Protein amino-terminal acetylation has potential effects on cell ...

show abstract

Section: Broad Substrate Specificity Of Nate May Account For Majoritysupporting

confidence: 58%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Human Naa50 Protein Displays Broad Substrate Specificity for Amino-terminal Acetylation

Reddi¹,

Saddanapu²,

Chinthapalli

et al. 2016

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…The majority of proteins undergo (cotranslational) N-terminal methionine excision (NME) by the action of methionine aminopeptidases in cytosol, plastids, and mitochondria. NME occurs if the Met is followed by a small residue (Gly, Ala, Thr, Pro, Ser, Val, and to a lesser degree Cys), which is referred to as the NME rule (Bonissone et al, 2013;Giglione et al, 2015). The NME rule can be explained in terms of steric hindrance of the methionine aminopeptidases action by the size of the side chain of the residue in the penultimate position.…”

Section: Protein Preprocessing Maturation and Different Proteoformsmentioning

confidence: 99%

Update: Post-translational protein modifications in plant metabolism

2015

View full text Add to dashboard Cite

ORCID ID: 0000-0001-9536-0487 (K.J.v.W.).Posttranslational modifications (PTMs) of proteins greatly expand proteome diversity, increase functionality, and allow for rapid responses, all at relatively low costs for the cell. PTMs play key roles in plants through their impact on signaling, gene expression, protein stability and interactions, and enzyme kinetics. Following a brief discussion of the experimental and bioinformatics challenges of PTM identification, localization, and quantification (occupancy), a concise overview is provided of the major PTMs and their (potential) functional consequences in plants, with emphasis on plant metabolism. Classic examples that illustrate the regulation of plant metabolic enzymes and pathways by PTMs and their cross talk are summarized. Recent large-scale proteomics studies mapped many PTMs to a wide range of metabolic functions. Unraveling of the PTM code, i.e. a predictive understanding of the (combinatorial) consequences of PTMs, is needed to convert this growing wealth of data into an understanding of plant metabolic regulation.The primary amino acid sequence of proteins is defined by the translated mRNA, often followed by Nor C-terminal cleavages for preprocessing, maturation, and/or activation. Proteins can undergo further reversible or irreversible posttranslational modifications (PTMs) of specific amino acid residues. Proteins are directly responsible for the production of plant metabolites because they act as enzymes or as regulators of enzymes. Ultimately, most proteins in a plant cell can affect plant metabolism (e.g. through effects on plant gene expression, cell fate and development, structural support, transport, etc.). Many metabolic enzymes and their regulators undergo a variety of PTMs, possibly resulting in changes in oligomeric state, stabilization/degradation, and (de)activation (Huber and Hardin, 2004), and PTMs can facilitate the optimization of metabolic flux. However, the direct in vivo consequence of a PTM on a metabolic enzyme or pathway is frequently not very clear, in part because it requires measurements of input and output of the reactions, including flux through the enzyme or pathway. This Update will start out with a short overview on the major PTMs observed for each amino acid residue (Table I), followed by a brief discussion of techniques for the characterization of protein PTMs, including determination of the localization within proteins (i.e. the specific residues) and occupancy. Challenges in dealing with multiple PTMs per protein and cross talk between PTMs will be briefly outlined. We then describe the major physiological PTMs observed in plants as well as PTMs that are nonenzymatically induced during sample preparation (Table II). Finally, this Update is completed with a number of well-studied, classical examples of the regulation of plant metabolism by PTMs, in particular for enzymes in primary metabolism (Calvin cycle, glycolysis, and respiration) and the C4 shuttle accommodating photosynthesis in C4 plants (Table III). As will be ev...

show abstract

Investigating Peptide‐Coenzyme A Conjugates as Chemical Probes for Proteomic Profiling of N‐Terminal and Lysine Acetyltransferases

et al. 2022

View full text Add to dashboard Cite

Acetyl groups are transferred from acetyl-coenzyme A (Ac-CoA) to protein N-termini and lysine side chains by N-terminal acetyltransferases (NATs) and lysine acetyltransferases (KATs), respectively. Building on lysine-CoA conjugates as KAT probes, we have synthesized peptide probes with CoA conjugated to Nterminal alanine (α-Ala-CoA), proline (α-Pro-CoA) or tri-glutamic acid (α-3Glu-CoA) units for interactome profiling of NAT complexes. The α-Ala-CoA probe enriched the majority of NAT catalytic and auxiliary subunits, while a lysine CoA-conjugate bound only a subset of endogenous KATs. Interactome profiling with the α-Pro-CoA probe showed reduced NAT recruitment in favor of metabolic CoA binding proteins and α-3Glu-CoA steered the interactome towards NAA80 and NatB. These findings agreed with the inherent substrate specificities of the target proteins and showed that N-terminal CoA-conjugated peptides are versatile probes for NAT complex profiling in lysates of physiological and pathological backgrounds.

show abstract

N-terminal protein modifications: Bringing back into play the ribosome

Cited by 159 publications

References 173 publications

Human Naa50 Protein Displays Broad Substrate Specificity for Amino-terminal Acetylation

Human Naa50 Protein Displays Broad Substrate Specificity for Amino-terminal Acetylation

Update: Post-translational protein modifications in plant metabolism

Investigating Peptide‐Coenzyme A Conjugates as Chemical Probes for Proteomic Profiling of N‐Terminal and Lysine Acetyltransferases

Contact Info

Product

Resources

About