Fact-checked by Grok 2 weeks ago

Pyruvate carboxylase

Pyruvate carboxylase (PC; EC 6.4.1.1) is a biotin-dependent mitochondrial enzyme that catalyzes the carboxylation of pyruvate to oxaloacetate using bicarbonate (HCO₃⁻) as the carboxyl donor and MgATP as the energy source, a reaction first described in 1959.^[1] This anaplerotic process is essential for replenishing intermediates in the tricarboxylic acid (TCA) cycle, thereby supporting various metabolic pathways including gluconeogenesis, lipogenesis, and amino acid synthesis.^[1] Structurally, PC is a homotetrameric protein (α₄) with each subunit approximately 120–130 kDa, comprising four functional domains: the biotin carboxylase (BC) domain, the biotin carboxyl carrier protein (BCCP) domain, the carboxyltransferase (CT) domain, and a unique allosteric regulatory domain.^[1] Biotin is covalently attached to a specific lysine residue within the BCCP domain, approximately 35 residues from the C-terminus of each subunit.^[1] The catalytic mechanism proceeds in two partial reactions: first, the BC domain uses ATP and bicarbonate to carboxylate the biotin prosthetic group, forming carboxybiotin and ADP plus inorganic phosphate; second, the CT domain transfers the carboxyl moiety from carboxybiotin to pyruvate, yielding oxaloacetate and regenerating free biotin.^[1] This process requires a divalent metal ion, such as Mg²⁺ or Mn²⁺, in the BC active site and a transition metal like Zn²⁺ in the CT site.^[1] PC activity is tightly regulated to match cellular metabolic demands, primarily through allosteric mechanisms: it is activated by acetyl-CoA, which promotes tetramer assembly, enhances biotin carboxylation, and increases substrate affinity, while L-aspartate serves as an inhibitor by stabilizing a less active conformation.^[1] Transcriptional regulation further modulates expression via factors such as peroxisome proliferator-activated receptor γ (PPARγ) in adipose tissue and cAMP response element-binding protein (CREB) in pancreatic β-cells.^[1] Physiologically, PC is vital in gluconeogenic tissues like the liver and kidney, where it provides oxaloacetate for glucose synthesis; in adipose tissue, it supports fatty acid production; and in pancreatic β-cells and astrocytes, it contributes to insulin secretion and neurotransmitter synthesis, respectively.^[1] Deficiency in PC, with an estimated incidence of 1 in 250,000 births, leads to severe metabolic disorders characterized by lactic acidosis, hyperammonemia, and developmental delays, underscoring its indispensable role in human metabolism.^[1]

Structure

Quaternary Assembly

Pyruvate carboxylase (PC) is a homotetrameric enzyme composed of four identical subunits, with a total molecular weight of approximately 500 kDa in mammals.^[2] Each monomer is a single polypeptide chain of about 1200 amino acids, typically around 1178-1300 residues depending on the species, enabling the enzyme's multifunctional catalytic capabilities.^[3]^[4] The tetrameric assembly is essential for activity, as isolated monomers or dimers lack full enzymatic function.^[4] The quaternary structure arranges the four subunits into a symmetric, square-shaped tetramer measuring roughly 180 × 120 × 140 Å, organized as two antiparallel dimers stacked into layers of dimers.^[4] Cryo-electron microscopy (cryo-EM) studies from 2010 revealed this architecture in detail, showing a central solvent channel that traverses the tetramer and positions the active sites peripherally on the monomer surfaces. The inter-subunit interfaces involve extensive contacts between the biotin carboxylase (BC), carboxyltransferase (CT), and pyruvate-binding (PT) domains across layers, such as BC-BC and PT-PT interactions, which stabilize the overall structure and facilitate long-range communication between subunits. These interfaces play a critical role in promoting cooperative function, allowing conformational changes in one subunit to influence others, which is vital for the enzyme's allosteric regulation and sequential catalysis. The tetrameric form is evolutionarily conserved across eukaryotes and most prokaryotes, underscoring its fundamental importance in metabolic pathways, though some archaea and bacteria exhibit α₄β₄ heterooctameric variants.^[4] This conservation highlights the structural adaptations that ensure stability and efficiency in diverse organisms.

Domains and Cofactors

Pyruvate carboxylase (PC) consists of four principal domains per monomeric subunit: the biotin carboxylase (BC) domain at the N-terminus, the carboxyltransferase (CT) domain, the biotin carboxyl carrier protein (BCCP) domain, and a central allosteric domain.^[5] The BC domain adopts an ATP-grasp fold typical of nucleotide-binding enzymes, while the CT domain features a two-domain architecture resembling other carboxyltransferases.^[6] The BCCP domain, located toward the C-terminus, functions as a flexible linker harboring the biotin prosthetic group.^[7] The allosteric domain, positioned between the BC and CT domains, is distinctive to PC and forms part of the enzyme's regulatory scaffold.^[5] Cofactor binding sites are distributed across these domains to support the enzyme's modular function. Biotin is covalently attached to a conserved lysine residue in the BCCP domain via an amide bond, enabling its role as a mobile carboxyl carrier.^[6] In the BC domain, ATP coordinates with Mg²⁺ ions at a site involving conserved motifs for phosphate and ribose binding.^[7] The CT domain accommodates pyruvate through interactions with its active site cleft and facilitates CO₂ incorporation via the incoming carboxybiotin from the BCCP.^[5] High-resolution structures have elucidated the structural organization of these domains. The full-length structure of PC from Rhizobium etli was determined at 2.0 Å resolution by X-ray crystallography in 2007, highlighting the compact arrangement of the BC, BCCP, CT, and allosteric domains within each monomer.^[6] For human PC, the C-terminal portion (encompassing the CT, BCCP, and allosteric domains) was resolved at 2.8 Å by X-ray crystallography in 2008, demonstrating the elongated, flexible BCCP "arm" that extends up to 80 Å to bridge distant active sites.^[5] More recent cryo-EM structures of full-length human PC, such as those at ~3.4 Å resolution in 2022 (PDB: 7WTE) and high-resolution in 2024 (PDB: 8HWL), confirm the domain organization, tetrameric assembly, and conformational dynamics in the human enzyme.^[8]^[9] These structures confirm the monomeric domains' independence, with the BCCP's mobility arising from a flexible linker region adjacent to the biotinylated lysine.^[7] The CT domain's active site includes a metal-binding motif that coordinates essential divalent cations. In R. etli PC, both Mn²⁺ and Zn²⁺ were identified at this site, where they polarize the pyruvate substrate's carbonyl group through direct ligation.^[6] The allosteric domain features a shallow binding pocket for acetyl-CoA, located at the N-terminal helix interfacing with the BC domain, which structurally stabilizes the monomer's conformation.^[5] These domain-specific features underpin PC's tetrameric assembly, where monomers interact primarily through their BC and allosteric domains.^[7]

Reaction Mechanism

Biotin Involvement

Pyruvate carboxylase (PC) utilizes biotin as a covalently bound prosthetic group, attached via an amide linkage to a specific lysine residue within the biotin carboxyl carrier protein (BCCP) domain. This post-translational modification, catalyzed by biotin protein ligase (also known as holocarboxylase synthetase), transforms the apo form of the enzyme into its active holoenzyme, enabling biotin to serve as a mobile carboxyl carrier during catalysis.^[10]^[11] The essential role of biotin in carboxylase enzymes was established in the 1950s, when studies demonstrated its requirement for CO₂ fixation reactions, such as the incorporation of ¹³CO₂ into oxaloacetate in cell-free extracts, marking biotin as a vitamin critical for these metabolic processes. This discovery built on earlier identifications of biotin deficiency symptoms and solidified its classification as a coenzyme for multiple biotin-dependent carboxylases, including PC.^[12]^[13] The biotinylated lysine residue exhibits remarkable conformational flexibility, functioning as a swinging arm that translocates the carboxybiotin moiety between the biotin carboxylase (BC) and carboxyl transferase (CT) active sites, spanning approximately 70 Å to facilitate carboxyl group transfer. This dynamic movement, supported by the flexible linker in the BCCP domain, ensures efficient coupling of the two partial reactions without requiring large-scale domain rearrangements.^[14] Biotin's structural features, particularly its ureido ring, confer specificity for CO₂ transfer in PC, allowing the formation of a stable carboxybiotin intermediate that delivers the carboxyl group to pyruvate without spontaneous hydrolysis, in contrast to other potential carriers like phosphoenolpyruvate that may release CO₂ more readily. This property underscores biotin's evolution as an optimal cofactor for precise, energy-efficient carboxylation in eukaryotic enzymes.^[15]^[16] Biotin-dependent carboxylases, including PC, trace their evolutionary origins to ancient metabolic pathways predating the last universal common ancestor, as evidenced by their widespread distribution across bacteria, archaea, and eukaryotes, reflecting an early adaptation for CO₂ assimilation in primordial autotrophy and carbon fixation processes.^[17]

Catalytic Steps

The catalytic mechanism of pyruvate carboxylase consists of two sequential partial reactions that together achieve the carboxylation of pyruvate to oxaloacetate. The overall reaction is:

\text{pyruvate} + \ce{HCO3-} + \ce{ATP} \to \text{oxaloacetate} + \ce{ADP} + \ce{P_i} + \ce{H+}

This process requires magnesium ions for ATP binding and is biotin-dependent, with the enzyme's prosthetic biotin group serving as a mobile carboxyl carrier between active sites.^[18] In the first partial reaction, occurring at the biotin carboxylase (BC) domain, bicarbonate is activated by ATP hydrolysis to form carboxybiotin. Specifically, bicarbonate, facilitated by a conserved lysine residue that enhances its nucleophilicity, attacks the γ-phosphorus of MgATP, yielding carboxyphosphate as a transient intermediate before carboxyl transfer to biotin and release of ADP and inorganic phosphate. This step couples the energy from ATP hydrolysis to bicarbonate activation, preventing uncoupled ATP breakdown. The second partial reaction takes place at the carboxyl transferase (CT) domain, where the activated carboxyl group from carboxybiotin is transferred directly to the enol form of pyruvate, generating oxaloacetate and regenerating the free biotin. The biotin "swings" between domains to link these spatially separated sites, ensuring efficient carboxyl delivery without dissociation.^[18]^[19]^[1] The kinetics follow an ordered bi-bi mechanism with ping-pong characteristics, reflecting the sequential binding of substrates and the intermediate biotin carboxylation step. Apparent Km values vary by species and conditions; for example, in chicken liver PC, they are approximately 0.07 mM for pyruvate, 0.11 mM for ATP, and 0.07 mM for HCO3-. These parameters highlight the enzyme's adaptation for physiological substrate concentrations in metabolic contexts like gluconeogenesis. Isotope exchange studies, using labeled pyruvate and oxaloacetate, have demonstrated rapid exchange without requiring other substrates, confirming the direct carboxyl transfer via the stable carboxybiotin intermediate and ruling out free CO2 or other diffusible species as obligatory intermediates.^[18]^[19]

Biological Functions

Gluconeogenesis and Anaplerosis

Pyruvate carboxylase (PC) functions as a critical anaplerotic enzyme in cellular metabolism by catalyzing the ATP-dependent carboxylation of pyruvate to form oxaloacetate within the mitochondria. This reaction replenishes tricarboxylic acid (TCA) cycle intermediates that are depleted during catabolic conditions, such as when α-ketoglutarate or oxaloacetate is diverted for amino acid synthesis or other biosynthetic pathways. By maintaining TCA cycle integrity, PC ensures sustained production of reducing equivalents and energy, preventing metabolic imbalances that could impair cellular function.^[20]^[21] In gluconeogenesis, PC plays an indispensable role by generating the oxaloacetate substrate required for phosphoenolpyruvate carboxykinase (PEPCK) to produce phosphoenolpyruvate, the first committed step bypassing the irreversible pyruvate kinase reaction of glycolysis. This process is particularly vital in the liver and kidney during fasting, where PC enables the synthesis of glucose from precursors like lactate and alanine to maintain blood glucose homeostasis. As a rate-limiting enzyme, PC controls flux through the gluconeogenic pathway, with over 80% of mitochondrial pyruvate being carboxylated in gluconeogenic tissues, directing a major portion of lactate-derived carbon toward glucose production in mammals.^[22]^[23]^[24] The oxaloacetate formed by PC cannot directly cross the mitochondrial inner membrane; instead, it integrates with the malate-aspartate shuttle for export to the cytosol. Within the mitochondria, oxaloacetate is reduced to malate by mitochondrial malate dehydrogenase, using NADH generated from the TCA cycle. Malate is then transported to the cytosol via the malate-α-ketoglutarate antiporter, where it is oxidized back to oxaloacetate by cytosolic malate dehydrogenase, providing NADH for gluconeogenic reactions and completing the shuttle. This mechanism ensures efficient transfer of reducing power and carbon skeletons, linking mitochondrial PC activity to cytosolic gluconeogenesis.^[25] In humans, PC activity is quantitatively linked to hepatic glucose output, serving as a key determinant of endogenous glucose production during fasting. Studies in type 2 diabetes models demonstrate that elevated PC flux contributes to excessive hepatic gluconeogenesis, while targeted inhibition of PC reduces plasma glucose levels and basal hepatic glucose production without altering insulin sensitivity, highlighting its therapeutic potential in metabolic disorders.^[26]

Tissue Distribution and Isoforms

Pyruvate carboxylase (PC) exhibits tissue-specific expression patterns that align with its roles in metabolic regulation, with highest levels observed in gluconeogenic organs such as the liver and kidney cortex, where it supports replenishment of tricarboxylic acid (TCA) cycle intermediates during carbohydrate limitation.^[1] Appreciable expression also occurs in adipose tissue and lactating mammary glands, reflecting adaptations for lipid metabolism and milk production, while levels are notably lower in skeletal muscle and brain, tissues that rely less on gluconeogenesis.^[1] In eukaryotes, PC is localized to the mitochondrial matrix, facilitated by an N-terminal mitochondrial targeting sequence that directs the protein for import and subsequent cleavage upon entry.^[27] In mammals, PC is encoded by a single gene, designated PC, located on chromosome 11q13.2 in humans, which produces multiple transcript variants through alternative promoter usage and splicing, resulting in tissue-specific transcript variants.^[28] For instance, the proximal promoter (P1) predominates in liver and adipose tissue, driving expression suited to gluconeogenic and lipogenic demands, whereas the distal promoter (P2) is active in brain, yielding transcripts with distinct regulatory elements in their 5' untranslated regions.^[1] In the brain, PC is predominantly expressed in astrocytes, where it functions as an anaplerotic enzyme to replenish TCA cycle intermediates, supporting the synthesis of neurotransmitter precursors such as glutamine for glutamate and GABA production.^[29] Similarly, in pancreatic β-cells, PC expression, regulated by the distal promoter, plays a crucial role in glucose-stimulated insulin secretion by providing oxaloacetate for anaplerosis and generating metabolic coupling factors like NADPH and ATP.^[30] In prokaryotes, PC variants diverge evolutionarily to fulfill anaplerotic functions in diverse contexts, such as sustaining TCA cycle flux during fermentation in bacteria like those in the genus Propionibacterium, where it enables oxaloacetate formation from pyruvate to support succinate production.^[31] Tissue-specific adaptations highlight PC's versatility; in adipocytes, it generates mitochondrial oxaloacetate to condense with acetyl-CoA, forming citrate that is exported to the cytosol for fatty acid synthesis via the citrate cleavage pathway, thereby linking carbohydrate metabolism to lipogenesis.^[1] Expression of PC is developmentally regulated, with upregulation in liver and kidney during fasting to enhance gluconeogenesis and in mammary glands during lactation to meet energy demands for milk synthesis.^[1] This dynamic control ensures metabolic flexibility across physiological states.

Regulation

Allosteric Control

Pyruvate carboxylase (PC) is subject to allosteric regulation primarily through the binding of acetyl-CoA, which acts as a potent activator in most species. Acetyl-CoA binds to a dedicated allosteric site within the central regulatory domain of the enzyme, stabilizing a catalytically competent conformation that enhances overall activity. This binding induces conformational changes that promote interdomain communication, particularly facilitating the efficient transfer of the carboxylated biotin intermediate between catalytic sites.^[32]^[1] The activation by acetyl-CoA significantly boosts enzymatic efficiency, increasing the maximum velocity (V_max) by approximately 10-fold in mammalian and certain bacterial forms, with a dissociation constant (K_d) around 10–50 μM. This effect is achieved by lowering the K_m for bicarbonate and accelerating the biotin carboxylation step, thereby linking PC activity to cellular energy status and metabolite availability. In contrast, ADP serves as a competitive inhibitor with ATP at the biotin carboxylase domain and reduces substrate affinity, which is particularly relevant in energy-depleted states where elevated ADP/ATP ratios signal low cellular energy.^[33]^[34]^[35] Species-specific variations in allosteric control reflect adaptations to metabolic environments; mammalian PC exhibits high sensitivity to acetyl-CoA activation, coordinating anaplerotic flux with tricarboxylic acid (TCA) cycle demands during nutrient shifts. In bacteria like Lactococcus lactis, PC is instead inhibited by the second messenger cyclic di-AMP (c-di-AMP), which binds at the carboxyltransferase dimer interface, inducing inhibitory conformational changes as detailed in a 2017 structural study.^[36]^[37] Structurally, acetyl-CoA binding promotes domain closure in the tetrameric enzyme, rigidifying the structure and enhancing the "biotin swing" mechanism where the carboxyl carrier protein shuttles between active sites more effectively. Physiologically, this allosteric activation by acetyl-CoA—derived from fatty acid β-oxidation—prevents futile cycling in the fed state by coupling oxaloacetate production to lipid catabolism, ensuring efficient gluconeogenesis without wasteful ATP hydrolysis. Recent studies (as of 2025) have identified small-molecule inhibitors targeting these allosteric sites, offering potential therapeutic insights into metabolic regulation.^[38]^[1]^[39]

Transcriptional and Post-Translational

The expression of the pyruvate carboxylase (PC) gene is primarily regulated at the transcriptional level to adapt to nutritional and hormonal cues. In hepatic tissue, glucagon elevates intracellular cAMP levels, activating protein kinase A and subsequent phosphorylation of CREB at Ser133, which binds to cAMP-responsive elements in the PC promoter to induce transcription during fasting or starvation. This mechanism ensures increased PC levels to support gluconeogenesis when glucose demand rises. Studies using transgenic mice expressing a dominant-negative CREB mutant demonstrated a significant reduction in hepatic PC mRNA, underscoring CREB's essential role in this induction. Conversely, insulin represses PC gene expression by selectively inhibiting the proximal promoter (P1), thereby suppressing gluconeogenic capacity in the fed state; this occurs through insulin signaling pathways that reduce transcription from this promoter, as observed in rat models where insulin treatment lowered PC mRNA in insulin-secreting cells and liver. Hormonal influences further modulate PC transcription, with upregulation observed in conditions like diabetes and starvation to enhance anaplerotic flux. The PC promoter contains glucocorticoid response elements that mediate induction by glucocorticoids, such as cortisol, which bind the glucocorticoid receptor to drive transcription and promote gluconeogenesis during stress or prolonged fasting. In diabetic states, elevated glucocorticoids and reduced insulin signaling amplify this effect, leading to higher PC activity and contributing to hyperglycemia. Post-translationally, PC undergoes proteolytic processing essential for its maturation and localization. The nascent PC polypeptide includes an N-terminal mitochondrial targeting sequence (approximately 30-40 amino acids) that directs import into the mitochondrial matrix via the TOM/TIM complex; upon translocation, this presequence is cleaved by mitochondrial processing peptidase to yield the mature 125 kDa enzyme, enabling full catalytic function. Emerging research as of 2025 highlights PC's role in immune cell regulation, such as in macrophages where it influences inflammatory responses in atherosclerosis through metabolic reprogramming. These long-term regulatory layers complement immediate allosteric activation by acetyl-CoA to maintain metabolic homeostasis.^[40]

Clinical Significance

Deficiency Disorders

Pyruvate carboxylase deficiency (PCD) is a rare autosomal recessive neurometabolic disorder caused by biallelic pathogenic variants in the PC gene on chromosome 11q13, leading to impaired conversion of pyruvate to oxaloacetate and subsequent disruptions in gluconeogenesis, anaplerosis, and energy metabolism.^[41] The condition is classified into three main types based on clinical severity: type A (infantile or North American form), type B (severe neonatal or French form), and type C (rare intermittent or attenuated form).^[41] Type A typically presents in infancy with moderate lactic acidosis, developmental delay, and failure to thrive, often allowing survival into early childhood with supportive care, while type B manifests neonatally with profound lactic acidemia, hyperammonemia, hypotonia, seizures, and respiratory distress, usually proving fatal within months without intervention.^[41] Type C is milder, with episodic decompensation and potential survival into adulthood, though it remains exceptionally rare with approximately 15 reported cases as of 2024.^[41] More than 100 pathogenic or likely pathogenic variants in the PC gene have been reported in ClinVar associated with PCD, including missense mutations, deletions, and splice site alterations that reduce enzyme activity to less than 10% of normal levels. Notable examples include the founder missense variant p.Ala610Thr, prevalent in North American Indigenous populations and associated with type A, and compound heterozygous mutations such as p.R631Q with other changes in type B cases, which disrupt biotin binding or catalytic domains.^[41]^[43] The estimated worldwide incidence is approximately 1 in 250,000 births, though it is higher in certain consanguineous or isolated communities, with approximately 75 cases documented globally as of 2024.^[44]^[41] Common symptoms across types include lactic acidemia, hypoglycemia, hyperammonemia, vomiting, lethargy, and neurological issues like hypotonia, ataxia, and epilepsy, stemming from accumulated toxic metabolites and energy deficits in brain and liver tissues.^[41] In type B, rapid progression to coma and multiorgan failure is typical, whereas type A may show intermittent metabolic crises triggered by fasting or illness.^[41] Diagnosis involves clinical suspicion prompted by neonatal screening (e.g., elevated citrulline), followed by confirmation via enzyme activity assay in fibroblasts or leukocytes showing reduced pyruvate carboxylation (<10% normal), and molecular genetic testing to identify PC variants.^[41] Differential diagnoses include other mitochondrial disorders like pyruvate dehydrogenase deficiency, ruled out by specific biochemical profiles.^[41] Treatment is primarily supportive, focusing on acute management with intravenous glucose, bicarbonate for acidosis, and hydration to stabilize metabolism during crises.^[41] Dichloroacetate has been used to activate pyruvate dehydrogenase and reduce lactate accumulation, though evidence is limited and primarily from small case series.^[45] High-dose biotin supplementation is often attempted but ineffective in most cases, as PCD is typically biotin-unresponsive, unlike holocarboxylase synthetase deficiency.^[45] Aspartate or citrate may provide alternative anaplerotic substrates, and thiamine supplementation supports related pathways, but outcomes vary.^[41] In severe type B cases, orthotopic liver transplantation has shown promise, reversing ketoacidosis, improving lactic acidemia, and enabling neurodevelopmental progress in survivors aged 6 months to 2.5 years at transplant.^[46] PCD was first described in 1968, with the severe neonatal (type B) form initially reported in French patients in the early 1970s, highlighting its French variant prevalence among certain populations.^[41] Early characterizations in the 1970s established the metabolic hallmarks, paving the way for genetic insights in subsequent decades.^[47]

Implications in Cancer and Metabolism

Pyruvate carboxylase (PC) plays a pivotal role in cancer progression, particularly in hypoxic tumor environments where it is upregulated to maintain redox balance and support biosynthetic demands.^[48] In glioblastoma stem cells, PC is essential for survival under hypoxia by facilitating pyruvate carboxylation, which replenishes oxaloacetate for the tricarboxylic acid cycle and enables aspartate synthesis critical for nucleotide production and cellular proliferation.^[49] This upregulation allows tumor cells to adapt to nutrient-limited, oxygen-poor niches, sustaining redox homeostasis via the malate-aspartate shuttle and mitigating oxidative stress. Studies have shown that PC expression is notably elevated in glioblastoma tissues compared to normal brain, underscoring its contribution to tumor aggressiveness.^[50] Inhibition of PC has been linked to suppressed cancer cell proliferation, as demonstrated in research by Sellers et al. (2021) highlighting its necessity for metabolic reprogramming in various malignancies.^[48] For instance, PC knockdown in non-small cell lung cancer models reduces growth by disrupting anaplerotic flux, limiting aspartate availability for protein and nucleotide synthesis.^[51] Similarly, in breast cancer, PC inhibition impairs metastasis by altering glucose and fatty acid utilization, emphasizing its therapeutic vulnerability in hypoxic settings.^[52] Beyond cancer, PC dysregulation contributes to metabolic diseases such as non-alcoholic fatty liver disease (NAFLD), where elevated hepatic PC flux promotes lipogenesis.^[53] Increased PC activity in NAFLD elevates oxaloacetate levels, driving citrate export from mitochondria to cytosol for fatty acid synthesis and exacerbating hepatic steatosis. In contrast, PC is downregulated in pancreatic islets of type 2 diabetes models, impairing pyruvate metabolism and gluconeogenesis, which disrupts insulin secretion and glucose homeostasis.^[54] Therapeutic strategies targeting PC hold promise for both cancer and metabolic disorders. PC inhibitors, including allosteric modulators like certain vitamin D analogs, have shown anti-cancer potential by inducing oxidative stress and halting proliferation in early breast cancer progression.^[55] Flux analysis in obesity models reveals PC as a key target, where its hepatic inhibition reduces gluconeogenesis, adiposity, and insulin resistance without severe hypoglycemia.^[56] Research from 2024 further implicates PC in tumor immune evasion via lactate modulation in the microenvironment; hypoxia-induced PC repression elevates lactate secretion from breast cancer cells, fostering an immunosuppressive milieu by recruiting pro-tumor macrophages and inhibiting T-cell function.^[57] Evolutionary adaptations of PC to fluctuating nutrient availability are dysregulated in modern high-carbohydrate diets, exacerbating metabolic syndrome through altered hepatic flux. In high-fat feeding paradigms mimicking contemporary Western diets, excessive PC activity sustains lipogenesis and gluconeogenesis, promoting insulin resistance and visceral fat accumulation characteristic of metabolic syndrome.^[56]

References

[1]
Structure, Mechanism and Regulation of Pyruvate Carboxylase - PMC
Pyruvate carboxylase (PC) is a biotin-containing enzyme that catalyses the HCO3−- and MgATP-dependent carboxylation of pyruvate to form oxaloacetate.
[2]
PC - Pyruvate carboxylase, mitochondrial - Homo sapiens (Human)
Oct 1, 1996 · Amino acids. 1178 (go to sequence). Protein existence. Evidence at ... EC number. EC:6.4.1.1 (UniProtKB | ENZYME | Rhea ) 1 publication.
[3]
Primary amino acid sequence and structure of human pyruvate ...
Oct 21, 1994 · The sequence of human PC has an open reading frame of 3537 nucleotides which encodes for a polypeptide with a length of 1178 amino acids.Missing: monomer | Show results with:monomer
[4]
https://www.cell.com/structure/fulltext/S0969-2126(10)00297-2
[5]
https://doi.org/10.1038/nsmb.1393
[6]
https://doi.org/10.1126/science.1144504
[7]
https://doi.org/10.1042/BJ20080709
[8]
Insight into the carboxyl transferase domain mechanism of pyruvate ...
In the first partial reaction, a bicarbonate-dependent ATP cleavage is coupled to the carboxylation of the covalently attached biotin prosthetic group in the ...
[9]
Domain Architecture of Pyruvate Carboxylase, a Biotin-Dependent ...
Aug 6, 2025 · Here we report the complete structure of pyruvate carboxylase at 2.0 angstroms resolution, which shows its domain arrangement. The structure, ...
[10]
[PDF] The role of cysteine 230 and lysine 238 of biotin carboxylase in the ...
In 1950, biotin again was found to be a necessary component of carboxylation reactions, the incorporation of 13CO2 into oxaloacetate in cell-free extracts of ...
[11]
Severo Ochoa, Harland Wood, and Feodor Lynen
May 27, 2004 · lonyl-CoA:pyruvate transcarboxylase, another biotin-dependent enzyme studied extensively by Harland Wood. Moreover, this organism had an ...
[12]
Structure and function of biotin-dependent carboxylases - PMC
Biotin-dependent carboxylases have two distinct enzymatic activities and catalyze their reactions in two steps [17, 18]. First, a biotin carboxylase (BC) ...Missing: 1950s | Show results with:1950s
[13]
Conformational Selection Governs Carrier Domain Positioning ... - NIH
Pyruvate carboxylase (PC) catalyzes the MgATP-dependent carboxylation of pyruvate, where the biotinylated carrier domain must translocate ~70 Å from the biotin ...
[14]
The enzymes of biotin dependent CO2 metabolism - PubMed Central
For example, if pyruvate is the acceptor then the enzyme is pyruvate carboxylase. Eukaryotic biotin-dependent enzymes only catalyze carboxylation reactions. In ...
[15]
A Substrate-induced Biotin Binding Pocket in the ...
Pyruvate carboxylase (PC), a multifunctional biotin-dependent enzyme, catalyzes the bicarbonate- and MgATP-dependent carboxylation of pyruvate to oxaloacetate, ...
[16]
Early evolution of the biotin-dependent carboxylase family - PMC
Biotin-dependent carboxylases are a diverse family of carboxylating enzymes widespread in the three domains of life, and thus thought to be very ancient.
[17]
Nearly 50 years in the making: defining the catalytic mechanism of ...
Jan 11, 2014 · Steady-state kinetic studies have examined the complex catalytic mechanism of the pyruvate carboxylase (PC).
[18]
https://febs.onlinelibrary.wiley.com/doi/10.1111/febs.12713
[19]
Anaplerosis of the citric acid cycle: role in energy ... - PubMed - NIH
... pyruvate carboxylase and propionyl-CoA carboxylase. Anaplerosis describes a pathway, which replenishes a metabolic cycle. We show that enzymes for ...
[20]
Anaplerotic roles of pyruvate carboxylase in mammalian tissues
In liver and kidney, PC provides oxaloacetate for gluconeogenesis. In adipocytes PC is involved in de novo fatty acid synthesis and glyceroneogenesis, and is ...
[21]
Structure, function and regulation of pyruvate carboxylase - PubMed
May 15, 1999 · In mammals, PC plays a crucial role in gluconeogenesis and lipogenesis, in the biosynthesis of neurotransmitter substances, and in glucose- ...
[22]
https://pubmed.ncbi.nlm.nih.gov/10229653/
[23]
Roles of pyruvate carboxylase in human diseases: from diabetes to ...
Pyruvate carboxylase (PC), an anaplerotic enzyme, plays an essential role in various cellular metabolic pathways including gluconeogenesis, de novo fatty acid ...
[24]
Biochemistry, Gluconeogenesis - StatPearls - NCBI Bookshelf - NIH
Jun 5, 2023 · Pyruvate carboxylation happens in mitochondria; then, via malate shuttle, oxaloacetate is being shuttled into the cytosol to be phosphorylated.
[25]
Targeting Pyruvate Carboxylase Reduces Gluconeogenesis ... - NIH
Pyruvate carboxylase catalyzes the first committed step for gluconeogenesis and is well poised to regulate hepatic glucose production. Pyruvate carboxylase is ...
[26]
Biochemical characterization, mitochondrial localization, expression ...
Removal of the protein's putative N-terminal mitochondrial targeting pre-sequence at the site of a predicted R-3 mitochondrial cleavage site (36 amino acids) ...
[27]
5091 - Gene ResultPC pyruvate carboxylase [ (human)] - NCBI
Sep 9, 2025 · Eight novel mutations were identified in PC from 8 pyruvate carboxylase deficiency patients. The crystal structures at 2.8-A resolution of ...
[28]
NADPH-generating systems in bacteria and archaea - PMC
A similar reaction sequence can involve PEP carboxylase (PEPCx) instead of pyruvate carboxylase, which also enables conversion of NADH into NADPH. This so ...
[29]
Mechanistic insight into allosteric activation of human pyruvate ...
Nov 3, 2022 · These structures and the biochemical studies reveal that acetyl-CoA stabilizes PC in a catalytically competent conformation, which triggers a cascade of events.Missing: Vmax Kd
[30]
Regulation of the structure and activity of pyruvate carboxylase ... - NIH
In this review we examine the effects of the allosteric activator, acetyl CoA on both the structure and catalytic activities of pyruvate carboxylase.
[31]
Effects of adenosine phosphates and nicotinamide nucleotides on ...
ADP, AMP, and adenosine inhibit pyruvate carboxylase. NADH also inhibits it. ADP and NADH effects are consistent with the [ATP]/[ADP] ratio.
[32]
Inhibitors of Pyruvate Carboxylase - PMC - NIH
Analogue of ADP, competitive inhibitor with respect to ATP. Enzyme has higher affinity for analogue as compared to ADP. Inhibition similar to ATP analogues.
[33]
Pyruvate carboxylase and cancer progression
Apr 30, 2021 · In nonmalignant tissue, PC plays an essential role in controlling whole-body energetics through regulation of gluconeogenesis in the liver ...
[34]
Structural and functional studies of pyruvate carboxylase regulation ...
Aug 14, 2017 · We report here structural, biochemical, and functional studies on the inhibition of Lactococcus lactis pyruvate carboxylase (LlPC) by c-di-AMP.
[35]
Allosteric regulation alters carrier domain translocation in pyruvate ...
Apr 11, 2018 · Pyruvate carboxylase (PC) catalyzes the ATP-dependent carboxylation of pyruvate to oxaloacetate. The reaction occurs in two separate ...
[36]
Pyruvate Carboxylase Deficiency - GeneReviews® - NCBI Bookshelf
Jun 2, 2009 · Pyruvate carboxylase (PC) deficiency is characterized in most affected individuals by failure to gain weight and/or linear growth failure, developmental delay, ...Diagnosis · Clinical Characteristics · Differential Diagnosis · Management
[37]
The Molecular Basis of Pyruvate Carboxylase Deficiency - NIH
Pyruvate carboxylase (PC) deficiency (OMIM, 266150) is a rare autosomal recessive disease. The revised PC gene structure described in this report consists of 20 ...
[38]
Genetics of Pyruvate Carboxylase Deficiency - Medscape Reference
Mar 15, 2019 · Pyruvate carboxylase deficiency is a rare disorder, with an approximate incidence of 1 in 250,000 births. Infantile-onset pyruvate carboxylase ...
[39]
Pyruvate Carboxylase Deficiency - Symptoms, Causes, Treatment
Pyruvate carboxylase deficiency (PC deficiency) is a rare genetic disorder present at or shortly after birth and characterized by failure to thrive.Missing: review | Show results with:review
[40]
Pyruvate carboxylase deficiency--insights from liver transplantation
Orthotopic hepatic transplantation completely reversed the ketoacidosis and the renal tubular abnormality and ameliorated the lactic acidemia.Missing: treatment | Show results with:treatment
[41]
Molecular Characterization of Pyruvate Carboxylase Deficiency in ...
May 1, 1998 · Crystal structures of human and Staphylococcus aureus pyruvate carboxylase and molecular insights into the carboxyltransfer reaction. Song ...