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N-terminus

The N-terminus, also known as the amino-terminus, is the end of a protein or polypeptide chain where the first amino acid residue possesses a free amino group (-NH₂, often protonated as -NH₃⁺ at physiological pH), marking it as the starting point of the linear sequence of amino acids linked by peptide bonds.^[1] This terminus contrasts with the C-terminus, which features a free carboxyl group (-COOH or -COO⁻), establishing the inherent polarity of proteins that dictates their directional synthesis during translation on ribosomes, proceeding from the N- to C-terminus in accordance with the 5' to 3' reading of mRNA.^[2] By convention, protein sequences are denoted from N- to C-terminus, with the N-terminal amino acid positioned at the left in structural representations.^[1] Beyond its structural role, the N-terminus plays critical functions in protein biogenesis and regulation, often harboring targeting signals that direct nascent polypeptides to specific subcellular locations, such as the endoplasmic reticulum for secretory proteins or mitochondria for organelle-resident enzymes.^[1] Post-translational modifications at the N-terminus, including the excision of the initiator methionine by methionine aminopeptidases (affecting over 50% of eukaryotic proteins), N-terminal acetylation by N-acetyltransferases (prevalent in 80-90% of human cytosolic proteins), and lipidations like myristoylation (occurring in ~0.5-3% of proteins), modulate protein stability, half-life, membrane association, and interactions with cellular machinery.^[3] These modifications enhance proteomic diversity and are essential for processes like signal transduction, protein degradation via the N-end rule pathway, and chromatin remodeling.^[3] In biochemistry, the N-terminus's sequence and modifications can profoundly influence higher-order protein folding, enzymatic activity, and disease associations; for instance, aberrant N-terminal processing is implicated in pathologies ranging from neurodegeneration to cancer,^[4] underscoring its biomedical significance.^[1]

Definition and Chemical Properties

Structural Definition

The N-terminus, also known as the amino-terminus, is the end of a protein or peptide chain where the alpha-amino group (-NH₂) of the first amino acid residue remains free and is not involved in a peptide bond, in contrast to the C-terminus where the carboxyl group is free.^[5] This structural feature defines the starting point of the polypeptide chain, with the N-terminal residue serving as the initial unit in the linear sequence of amino acids.^[6] At the atomic level, the N-terminus comprises the alpha-amino group, which is typically protonated to NH₃⁺ at physiological pH (around 7.4) due to its basic properties, the tetrahedral alpha-carbon atom bonded to a hydrogen, the side chain (R group) specific to the residue, and the carbonyl group that forms the first peptide bond with the subsequent residue.^[5] The general structural formula for the N-terminal residue is H₂N-CH(R)-CO-, where the amino group is at one end and the carbonyl links to the nitrogen of the next residue in the chain, forming the repeating backbone motif -NH-CH(R)-CO-.^[6] This configuration distinguishes the N-terminus as the nucleophilic end of the chain, with the alpha-amino nitrogen serving as a key reactive site in the absence of further bonding.^[5] In standard biochemical notation, protein sequences are written directionally from the N-terminus to the C-terminus, reflecting the biosynthetic order, with the N-terminal amino acid assigned position 1 and subsequent residues numbered sequentially.^[5] This convention facilitates unambiguous description of primary structure and is universally adopted in protein databases and literature.^[6] The terminology "N-terminus" emerged in the mid-20th century, building on early protein chemistry studies, particularly Frederick Sanger's pioneering sequencing of insulin in the 1940s and 1950s, which confirmed the N-to-C directionality of polypeptide chains through identification of N-terminal residues like phenylalanine and glycine.^[7] Sanger's development of methods to label and isolate N-terminal groups, such as using dinitrophenyl (DNP) derivatives, provided the experimental foundation for recognizing and naming these chain ends.^[8]

Chemical Reactivity and Properties

The alpha-amino group at the N-terminus of a protein typically exhibits a pKa value in the range of 9 to 10, depending on the specific amino acid and local environment.^[9] At physiological pH values around 7, this group is predominantly protonated, existing as a positively charged ammonium ion (NH₃⁺), which contributes a net positive charge to the overall protein structure.^[10] This protonation state arises because the pH is below the pKa, favoring the conjugate acid form according to the Henderson-Hasselbalch equation.^[11] The N-terminal amino group possesses nucleophilic character due to the availability of its lone pair on nitrogen, enabling it to participate in reactions such as the formation of Schiff bases (imines) with aldehydes under physiological conditions.^[12] This reactivity is notably higher than that of the nitrogen atoms within internal peptide bonds, as the latter are part of amide linkages where resonance stabilization delocalizes the lone pair into the adjacent carbonyl group, reducing nucleophilicity.^[13] For instance, the free alpha-amino group reacts more readily with electrophiles like carbonyl compounds compared to the less available amide nitrogens.^[14] The positive charge from the protonated N-terminus influences the isoelectric point (pI) of the protein, defined as the pH at which the net charge is zero, particularly in smaller peptides where terminal charges have a more pronounced effect relative to side-chain contributions.^[15] Acetylation of the N-terminus, for example, neutralizes this charge, shifting the pI to a lower value by removing the positive contribution.^[16] In contrast to the N-terminus, which imparts a basic character through its positive charge, the C-terminal carboxyl group has a pKa of approximately 2 to 3 and is deprotonated (COO⁻) at neutral pH, providing an acidic, negative charge that balances the protein's overall electrostatic properties.^[9] The reactivity of the N-terminal group is modulated by its solvent exposure within the protein structure; exposed termini in unstructured or surface regions exhibit higher accessibility to solvents and reagents, enhancing nucleophilic interactions, whereas buried N-termini in folded cores experience steric hindrance and hydrophobic shielding that diminish reactivity.^[17] This environmental dependence underscores the role of protein conformation in regulating chemical behavior at the N-terminus.^[18]

Biological Synthesis and Initial Processing

Role in Protein Translation

In prokaryotes, protein translation initiates with the aminoacylation of the initiator tRNA (tRNAfMet) by methionyl-tRNA synthetase to form Met-tRNAfMet, followed by formylation of the methionine residue by methionyl-tRNA formyltransferase, yielding N-formylmethionine-tRNAfMet (fMet-tRNAfMet). This modified initiator tRNA, along with mRNA and the 30S ribosomal subunit, forms the 30S pre-initiation complex in a process facilitated by initiation factors IF1, IF2, and IF3; IF2 specifically delivers the fMet-tRNAfMet to the P-site of the ribosome, ensuring the N-terminal residue is fMet. The resulting N-terminus of the nascent polypeptide is thus formylated methionine, which positions it at the amino end of the growing chain.^[19] In eukaryotes, translation initiation differs in that the initiator tRNA (tRNAiMet) is charged with unmodified methionine by methionyl-tRNA synthetase to form Met-tRNAiMet, without formylation. This Met-tRNAiMet forms a ternary complex with eukaryotic initiation factor 2 (eIF2) bound to GTP, which is then recruited to the 40S ribosomal subunit pre-assembled with mRNA via the 5' cap and scanning for the AUG start codon, aided by other factors like eIF1, eIF1A, eIF3, and eIF5. The eIF2-GTP-Met-tRNAiMet complex ensures precise delivery of the initiator to the P-site, establishing the N-terminal methionine as the first residue of the polypeptide. This mechanism maintains fidelity in N-terminal selection, preventing non-AUG starts.^[20]^[21] During elongation in both domains of life, the ribosome adds new amino acids to the carboxyl end of the growing chain via peptidyl transferase activity, resulting in vectorial synthesis from N- to C-terminus; the nascent polypeptide emerges from the ribosomal exit tunnel with the N-terminus first, approximately 30-40 residues ahead of the peptidyl-tRNA in the P-site. This N-to-C directionality is universal across all organisms, reflecting the chemical constraint of peptide bond formation where the α-amino group of the incoming aminoacyl-tRNA attacks the carbonyl carbon of the peptidyl-tRNA, extending the chain carboxyl-terminally. The evolutionary conservation of this directionality traces to the RNA world hypothesis, where primordial ribozymes likely catalyzed similar N-to-C peptide linkages, providing a foundational mechanism for coded protein synthesis that persisted through the transition to modern ribosomes.^[22]^[23]

N-Terminal Processing Mechanisms

In eukaryotes, the nascent N-terminus of newly synthesized proteins typically begins with an N-terminal methionine (Met) residue, which is often excised by methionine aminopeptidases (MetAPs) to generate the mature protein form.^[24] This N-terminal methionine excision (NME) occurs in approximately two-thirds of eukaryotic proteins and is catalyzed by two main enzymes: MAP1 and MAP2, which exhibit overlapping but distinct substrate specificities.^[24] Both MAP1 and MAP2 cleave the initiator Met when the second residue (P2 position) is alanine (Ala), cysteine (Cys), glycine (Gly), proline (Pro), serine (Ser), threonine (Thr), or valine (Val), though MAP2 shows higher efficiency for certain residues like Thr and Val.^[25] The efficiency of excision is thus heavily influenced by the identity of the residues at positions 1 (Met) and 2, with steric hindrance from larger side chains at P2 generally preventing cleavage.^[25] In prokaryotes, translation initiates with N-formylmethionine (fMet), and processing begins with the removal of the formyl group by peptide deformylase (PDF), an iron-dependent enzyme that acts cotranslationally on the nascent chain as it emerges from the ribosome.^[26] Following deformylation, the exposed Met is often further removed by prokaryotic MetAPs if the penultimate residue is small and uncharged, similar to the eukaryotic specificity rules, though prokaryotic MetAPs show variations in preference.^[27] This sequential processing ensures the rapid maturation of the N-terminus, with PDF's reversible association to the ribosomal exit tunnel facilitating efficient access to the fMet substrate.^[26] Signal peptide cleavage represents another key N-terminal processing mechanism, particularly for proteins destined for secretion or membrane insertion, where an N-terminal signal peptide (typically 15-30 residues) directs translocation across membranes and is subsequently removed by endoproteolytic cleavage.^[28] This cleavage is mediated by signal peptidases, such as the endoplasmic reticulum-resident signal peptidase complex (SPC) in eukaryotes or the simpler SecA/SecY-dependent peptidase in bacteria, which recognize a consensus cleavage site featuring small residues (e.g., Ala or Ser) at positions -1 and -3 relative to the scissile bond.^[28] The process exposes the mature N-terminus and is essential for proper protein folding and function post-translocation. N-terminal processing mechanisms are predominantly co-translational, occurring as the polypeptide emerges from the ribosome to minimize misfolding and aggregation risks, though some post-translational events can occur in the cytosol or organelles depending on chaperone availability and enzyme localization.^[27] For instance, PDF and MetAPs in prokaryotes associate directly with ribosomes for immediate action, while eukaryotic MAPs and signal peptidases often couple processing to ongoing translation or early translocation steps for efficiency.^[27] Factors such as the local cellular environment, including metal ion availability (e.g., cobalt or iron cofactors for MetAPs and PDF), and the rate of translation elongation further modulate the propensity and timing of these cleavages.^[29]

Functions in Cellular Localization

Signal Peptides for Secretion

Signal peptides are short amino acid sequences located at the N-terminus of nascent polypeptides that direct proteins into the secretory pathway for export from the cell or insertion into membranes. These peptides typically consist of 15-30 residues organized into three distinct regions: an N-region (1-5 residues) that is positively charged and hydrophilic, serving as an initial targeting signal; a central H-region (7-15 residues) forming a hydrophobic α-helix that interacts with the lipid bilayer or targeting machinery; and a C-region (3-7 residues) containing a cleavage site motif, often Ala-X-Ala (where X is any amino acid except proline), which allows for precise removal by signal peptidase.^[30] This tripartite structure ensures efficient recognition and translocation, with the hydrophobic core being critical for membrane partitioning during the targeting process. Upon emergence from the ribosome during co-translational synthesis, the signal peptide is recognized by the signal recognition particle (SRP), a ribonucleoprotein complex that binds the hydrophobic H-region, pauses translation, and facilitates docking to the SRP receptor on the endoplasmic reticulum (ER) membrane in eukaryotes or the SecYEG translocon in prokaryotes. This interaction initiates translocation through the Sec61 translocon in eukaryotes, where the polypeptide chain is threaded into the ER lumen, driven by the nascent chain and GTP hydrolysis. In prokaryotes, the process is analogous but occurs at the plasma membrane via the SecA ATPase for post-translational translocation in some cases. Following translocation, the signal peptide is cleaved by signal peptidase—a multi-subunit enzyme complex—exposing the mature N-terminus of the protein; this cleavage occurs at the Ala-X-Ala motif and is essential for proper folding and function of secreted proteins. Signal peptides are ubiquitous in the secretory pathway and are present in approximately 15% of human proteins, encompassing secreted proteins such as antibodies, hormones, and extracellular matrix components, where they are indispensable for directing these molecules out of the cell.^[31] In eukaryotes, they ensure targeting to the classical ER-Golgi-plasma membrane route, while in prokaryotes, they guide proteins to the periplasm or outer membrane. Variations between bacterial and eukaryotic signal peptides include differences in average length (22-23 residues in eukaryotes versus 24-26 in Gram-negative bacteria) and hydrophobicity of the H-region, with bacterial peptides often exhibiting greater hydrophobicity to accommodate the SecYEG channel; additionally, eukaryotic N-regions tend to be more positively charged to enforce the positive-inside rule for membrane orientation.^[32] These adaptations reflect evolutionary divergences in translocation machinery while maintaining the core functional principles.^[33]

Organelle Targeting Sequences

N-terminal targeting sequences direct nuclear-encoded proteins to intracellular organelles such as mitochondria, chloroplasts, and peroxisomes, distinguishing them from secretory signal peptides by facilitating import across organelle membranes rather than the endoplasmic reticulum. These sequences are typically cleavable presequences that ensure proper localization during protein biogenesis, with their recognition by specific receptors initiating translocation through dedicated translocon complexes.^[34] Mitochondrial targeting peptides (MTPs), also known as presequences, are N-terminal extensions of 20–80 amino acids that adopt an amphipathic α-helical structure, characterized by a positively charged face rich in arginine and lysine residues and a hydrophobic face. This amphipathicity enables interaction with the negatively charged inner mitochondrial membrane and recognition by receptors like Tom20. Upon import, MTPs are cleaved by the mitochondrial processing peptidase (MPP) in the matrix, releasing the mature protein; MPP recognizes specific cleavage motifs, often involving arginine at the -2 or -10 position relative to the cleavage site.^[35]^[34] Chloroplast transit peptides, found in nuclear-encoded proteins destined for plastids, are generally longer than MTPs, ranging from 40–70 residues on average (up to 146 in some cases), and exhibit less defined secondary structure with an enrichment in serine and threonine residues alongside hydroxylated amino acids. Unlike the helical MTPs, these peptides are often unstructured but form transient helices during import, facilitating passage through the TOC-TIC translocons. Processing occurs in the stroma by the stromal processing peptidase (SPP), which removes the transit peptide in one or two steps, depending on the precursor.^[34]^[36] Peroxisomal targeting signals include N-terminal variants like the PTS2 motif, a cleavable sequence typically consisting of (R/K)(L/V/I/Q)XX(L/V/I/H/Q)(L/A), located near the N-terminus after a short basic stretch; it is recognized by the receptor Pex7 and often processed by the peroxisomal processing peptidase (PEX1/PEX6). In contrast to the C-terminal PTS1 (e.g., SKL motif), PTS2 directs a subset of peroxisomal enzymes, such as those involved in fatty acid β-oxidation, and resembles other N-terminal signals in its bipartite structure with a targeting and cleavage domain.^[37]^[38] Import into mitochondria is mediated by the TOM-TIM supercomplex, where the TOM complex in the outer membrane acts as the general entry gate—featuring receptors Tom20 and Tom22 for presequence recognition and the β-barrel pore Tom40 for translocation—while the TIM23 complex in the inner membrane, including the channel Tim23 and receptor Tim50, drives matrix import in a membrane potential-dependent manner, often coupled with the presequence translocase-associated motor (PAM). Chloroplast import similarly relies on the TOC-TIC supercomplex spanning both envelope membranes, with TOC components like Toc34/Toc159 (receptors) and Toc75 (pore) initiating recognition of transit peptides, bridged to the inner TIC translocon (including Tic20 and Tic214 channels) via an intermembrane space scaffold for coordinated translocation powered by stromal ATP and GTP hydrolysis. Peroxisomal import of PTS2 proteins involves Pex7 receptor binding in the cytosol, docking to the peroxisomal membrane via Pex14/Pex13, and translocation through a transient pore without a stable translocon like TOM or TOC.^[39]^[40]^[41] The evolutionary origins of these N-terminal targeting sequences trace back to the endosymbiotic events that gave rise to mitochondria and chloroplasts from bacterial ancestors, with evidence suggesting that modern presequences evolved from antimicrobial peptides of the host cell, which initially targeted the endosymbiont genomes before gene transfer to the nucleus necessitated import signals. This antimicrobial heritage is reflected in their amphipathic, cationic properties that disrupt bacterial membranes, adapted over time for specific organelle recognition in eukaryotes. Peroxisomal signals may have arisen independently, possibly from ancient peroxisome-like compartments predating endosymbiosis.^[42]^[43]

Regulatory Roles

N-End Rule Pathway

The N-end rule pathway is a conserved proteolytic system that links the in vivo half-life of a protein to the identity of its N-terminal residue, thereby regulating protein stability and degradation. Discovered in the 1980s by Alexander Varshavsky and colleagues through studies on engineered substrates in Saccharomyces cerevisiae, the pathway was first described as a ubiquitin-dependent process where specific N-terminal amino acids act as degradation signals, or N-degrons.^[44] This principle, termed the N-end rule, posits that proteins bearing destabilizing N-terminal residues are rapidly degraded, while those with stabilizing residues persist longer, influencing cellular protein levels without relying on transcription or translation changes.^[45] The N-end rule organizes destabilizing residues into a hierarchy across primary, secondary, and tertiary levels in eukaryotes. Primary destabilizers include basic residues like Arg, Lys, and His (type 1), and bulky hydrophobic residues such as Phe, Leu, Ile, Trp, and Tyr (type 2), which are directly recognized and confer short half-lives of 2–30 minutes. Secondary destabilizers, Asp and Glu, are conditionally destabilizing and require post-translational arginylation by arginyl-tRNA-protein transferases (e.g., ATE1 in eukaryotes) to become primary-like Arg at the N-terminus. Tertiary destabilizers, Asn and Gln, are first deamidated to Asp and Glu, respectively, by N-terminal amidases (e.g., NTAN1 for Asn), followed by arginylation; additionally, Cys can become secondary via oxidation to Cys-sulfinate or Cys-sulfonate by oxygen or nitric oxide, enabling arginylation. In contrast, N-terminal Met, Gly, Ala, Ser, Thr, Val, and Pro are stabilizing, with half-lives exceeding 10 hours or more.^[45] This hierarchy often becomes relevant after initial N-terminal processing exposes the mature residue, such as through methionine excision.^[46] In eukaryotes, the mechanism involves recognition by E3 ubiquitin ligases known as N-recognins, primarily UBR1 (and UBR2 in mammals), which bind primary destabilizers via specialized domains like the UBR box and Clk-1 type zinc finger. This leads to polyubiquitination of the substrate via the E1-E2 cascade, targeting it for degradation by the 26S proteasome. Secondary and tertiary residues are converted stepwise to primary forms before recognition. In bacteria, the pathway is ubiquitin-independent; the adaptor ClpS binds primary destabilizers (e.g., Leu, Phe, Trp, Tyr) and delivers substrates to the ClpAP protease for ATP-dependent unfolding and degradation, with no equivalent hierarchy beyond primary residues. The pathway is evolutionarily conserved from prokaryotes to humans, adapting to diverse cellular needs.^[45] Biologically, the N-end rule pathway plays critical roles in development, such as cardiovascular patterning and neurogenesis via targeted degradation of transcription factors, and in stress responses, including hypoxia sensing through Cys oxidation and arginylation in plants and animals. It maintains protein quality control by eliminating misfolded or damaged proteins and regulates key processes like DNA repair and apoptosis. Although exact proteome coverage varies, the pathway influences a substantial fraction of cellular proteins, particularly those exposed to conditional N-degrons during stress or processing.^[45] The N-end rule has been expanded beyond the classical Arg/N-end rule to include additional N-degron pathways. The Ac/N-end rule pathway, where N-terminal acetylation (common in eukaryotes) acts as a degron recognized by Doa10 E3 ligase, regulates stability of ~10-20% of acetylated proteins, affecting processes like cell cycle and ER quality control. In yeast, a Pro/N-end rule pathway targets N-terminal proline via GID E3 ligase, degrading gluconeogenic enzymes during respiratory growth shifts. These extensions, identified as of 2017-2024, broaden the pathway's regulatory scope to over 30% of the proteome in some contexts.^[47]^[48]

Influence on Protein-Protein Interactions

The N-terminus of proteins often contains specific motifs that serve as binding sites for Src homology 3 (SH3) domains, enabling key protein-protein interactions in signaling pathways. These interactions position proteins in multi-protein scaffolds, facilitating downstream phosphorylation events in processes like cell adhesion and proliferation. Nascent polypeptides emerging from the ribosome expose their N-termini first, making them prime targets for chaperone interactions to ensure proper folding. The Hsp70 chaperone family, including the yeast homolog Ssb, binds to these exposed nascent chains near the ribosome exit tunnel, shielding hydrophobic segments and preventing aggregation or misfolding during cotranslational folding.^[49] This binding is selective for substrates with slow translation kinetics or intrinsic folding challenges, thereby promoting efficient maturation of proteins. The exposure or processing of the N-terminus can exert allosteric effects on enzymatic activity through conformational changes that alter interaction interfaces. In calpains, a family of calcium-dependent proteases, the N-terminal domain I undergoes autolytic cleavage upon activation, exposing residues that relieve autoinhibition and enhance substrate access to the catalytic core. This N-terminal remodeling modulates calpain's proteolytic activity in cytoskeletal dynamics and apoptosis.^[50] Pathological alterations in the N-terminus can disrupt protein-protein interactions, contributing to neurodegenerative diseases. In Alzheimer's disease, mutations in the N-terminal projection domain of tau protein, such as the R5L variant, impair tau's binding to partners like BIN1 and microtubules, promoting aberrant aggregation into neurofibrillary tangles.^[51] These changes underscore the N-terminus's critical role in maintaining interaction networks for neuronal integrity.

Post-Translational Modifications

N-Terminal Acetylation

N-terminal acetylation is the most common post-translational modification at the protein N-terminus in eukaryotes, affecting approximately 80% of the human proteome. This modification involves the transfer of an acetyl group from acetyl-coenzyme A (AcCoA) to the α-amino group of the N-terminal residue, catalyzed primarily by N-terminal acetyltransferase (NAT) complexes in a co-translational process on the ribosome. The reaction neutralizes the positive charge of the α-amino group, altering the protein's biophysical properties and influencing downstream cellular processes.^[52] The specificity of N-terminal acetylation is determined by the N-terminal amino acid sequence and is mediated by distinct NAT complexes. The NatA complex, composed of the catalytic subunit NAA10 and auxiliary subunit NAA15, targets proteins after methionine excision by methionine aminopeptidase, particularly those starting with serine, alanine, glycine, threonine, or cysteine, acetylating about 40% of eukaryotic proteins. NatB, formed by NAA20 (catalytic) and NAA25 (auxiliary), acetylates unprocessed N-termini beginning with methionine followed by charged residues such as aspartate, glutamate, or asparagine, accounting for roughly 17-20% of the proteome. NatC, involving NAA30, NAA35, and NAA38, modifies methionine-initiated sequences with hydrophobic second residues like leucine or isoleucine, while NatD (NAA40) is more restricted, acetylating histone H2A and H4 with N-terminal serine or alanine. These complexes ensure broad coverage of the proteome while exhibiting high substrate selectivity.^[52]^[53] The functional consequences of N-terminal acetylation are multifaceted, primarily affecting protein stability, folding, and degradation. By capping the N-terminus, acetylation often blocks the N-end rule pathway, which targets proteins with certain destabilizing N-terminal residues for ubiquitin-proteasome degradation, thereby promoting overall protein half-life and stability—for example, acetylated actin and tropomyosin exhibit extended lifespans compared to their non-acetylated counterparts. It also facilitates proper protein folding by influencing secondary structure formation; in the case of α-synuclein, acetylation enhances α-helical propensity and reduces aggregation propensity, mitigating neurotoxicity in Parkinson's disease models. Conversely, in specific contexts, N-terminal acetylation can create Ac/N-degrons that recruit E3 ligases like Not4 for targeted degradation, as observed with certain cytosolic proteins. These effects underscore acetylation's role as a regulatory switch in proteostasis.^[52]^[54]^[54] N-terminal acetylation is generally irreversible, with no dedicated N-terminal deacetylases (NDACs) identified to date, unlike the dynamic lysine acetylation regulated by histone deacetylases (HDACs) and sirtuins. This stability contributes to its persistent influence on protein function throughout the cellular lifetime.^[55] Dysregulation of N-terminal acetylation is linked to disease, particularly cancer, where aberrant NAT activity drives tumorigenesis. Overexpression of NAT10 (also known as NAA10), the catalytic subunit of NatA, is frequently observed in prostate, breast, and colorectal cancers, promoting cell proliferation, invasion, and metastasis through enhanced acetylation of oncogenic substrates like β-catenin stabilizers. For instance, elevated NAT10 levels correlate with poor prognosis in pancreatic ductal adenocarcinoma by stabilizing key regulatory proteins and suppressing ferroptosis. These findings highlight NAT10 as a potential therapeutic target in oncology.^[52]^[56]^[57]

Lipid and Acyl Modifications

Lipid and acyl modifications at the N-terminus of proteins involve the covalent attachment of fatty acids, enhancing hydrophobicity and facilitating membrane association. These irreversible or reversible modifications target specific residues, such as glycine or cysteine, and are crucial for protein localization, stability, and signaling functions in eukaryotic cells.^[58] N-myristoylation is an irreversible co-translational modification where myristic acid, a 14-carbon saturated fatty acid, is attached via an amide bond to the α-amino group of an N-terminal glycine residue. This process is catalyzed by N-myristoyltransferases (NMTs), ubiquitous enzymes in eukaryotes, with humans expressing NMT1 and NMT2 isoforms that share high sequence similarity and overlapping substrate specificities.^[58]^[59]^[60] For myristoylation to occur, the initiator methionine must first be removed by methionine aminopeptidases, exposing the glycine at position 2 in the consensus sequence MGXXX(S/T). This modification affects approximately 0.5% of eukaryotic proteins, corresponding to about 150 substrates in humans, and promotes weak but specific interactions with phospholipid bilayers.^[61] Representative examples include Src family kinases, where N-myristoylation anchors the SH4 domain to the plasma membrane, enabling kinase activation and signal transduction in pathways like cell growth and motility.^[62]^[63] Beyond membrane targeting, myristoylation influences protein conformation and stability, often cooperating with other modifications for full functionality.^[64] These N-terminal lipid modifications increase membrane affinity and enable precise spatiotemporal regulation of protein function, with myristoylation providing stable anchoring. Dysregulation of these processes is implicated in diseases such as cancer, where inhibitors targeting NMTs show therapeutic potential by disrupting oncogenic signaling.^[65]

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Dec 15, 2023 · This study was aimed to elucidate the high-resolution structures of human mitochondrial MetAP1D in its apo-, cobalt-, and methionine-bound states.
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The cleavage of the N-terminal 126 amino acid residues of pre-gp130Env containing the signal anchor sequence occurs during intracellular transport by a furin- ...
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Signal peptide discrimination and cleavage site identification using ...
Feb 1, 2014 · Almost 15% of human proteins contain SPs [3] and such proteins are either secreted or inserted into membranes as type I membrane proteins.
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Distribution of lengths of the eukaryotic and prokaryotic signal peptides. The average length is 22.6 amino acids for eukaryotes, 25.1 for Gram-negative ...
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In this review, we focus on characterizing the structure, function, and ... Cleavage-site motifs in mitochondrial targeting peptides. Protein Eng. Des ...Missing: paper | Show results with:paper
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Evolution and Design Principles of the Diverse Chloroplast Transit ...
The N-terminal cleavable transit peptide (TP) is necessary and sufficient for the import of nucleus-encoded interior chloroplast proteins.
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Structurally, PTS2 motifs resemble other N-terminal targeting signals, whereas the functional relation to the second peroxisomal targeting signal (PTS1) is ...
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To specify the C-terminal peroxisome targeting signal type 1 (PTS1) and the N-terminal PTS2 for higher plants, a maximum number of plant cDNAs and expressed ...
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Proline-rich sequences that bind to Src homology 3 domains ... - PNAS
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N-acetyltransferase 10 promotes colon cancer progression by ...
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Myristoylation is critical for membrane association of Src kinases, but a role for myristate in regulating other aspects of Src biology has not been explored.
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Protein N-myristoylation plays a critical role in the mitochondrial ...
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Dynamic palmitoylation events following T-cell receptor signaling
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Abstract. Protein palmitoylation describes the post-translational modification of cysteines by a thioester-linked long chain fatty acid.
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Protein lipidation in health and disease: molecular basis ... - Nature
Mar 15, 2024 · S-palmitoylation plays a role in regulating various cellular signal transduction pathways, primarily by influencing the aforementioned protein ...
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Jan 9, 2023 · This study demonstrates that desloratadine may be a novel anticancer drug and that NMT1-mediated myristoylation contributes to HCC progression.