PCKGC_PIG
ID PCKGC_PIG Reviewed; 622 AA.
AC A0A4X1UM84; A5GFS4; K7GND0;
DT 12-AUG-2020, integrated into UniProtKB/Swiss-Prot.
DT 12-AUG-2020, sequence version 2.
DT 03-AUG-2022, entry version 13.
DE RecName: Full=Phosphoenolpyruvate carboxykinase, cytosolic [GTP] {ECO:0000305};
DE Short=PEPCK-C;
DE EC=4.1.1.32 {ECO:0000250|UniProtKB:P35558};
DE AltName: Full=Serine-protein kinase PCK1 {ECO:0000305};
DE EC=2.7.11.- {ECO:0000250|UniProtKB:P35558};
GN Name=PCK1 {ECO:0000303|PubMed:26792594};
GN Synonyms=PEPCK1 {ECO:0000250|UniProtKB:P35558};
OS Sus scrofa (Pig).
OC Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
OC Eutheria; Laurasiatheria; Artiodactyla; Suina; Suidae; Sus.
OX NCBI_TaxID=9823;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RG Porcine genome sequencing project;
RL Submitted (MAY-2007) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, VARIANT
RP LEU-139, AND CHARACTERIZATION OF VARIANT LEU-139.
RX PubMed=26792594; DOI=10.1038/srep19617;
RA Latorre P., Burgos C., Hidalgo J., Varona L., Carrodeguas J.A.,
RA Lopez-Buesa P.;
RT "c.A2456C-substitution in Pck1 changes the enzyme kinetic and functional
RT properties modifying fat distribution in pigs.";
RL Sci. Rep. 6:19617-19617(2016).
CC -!- FUNCTION: Cytosolic phosphoenolpyruvate carboxykinase that catalyzes
CC the reversible decarboxylation and phosphorylation of oxaloacetate
CC (OAA) and acts as the rate-limiting enzyme in gluconeogenesis
CC (PubMed:26792594). Regulates cataplerosis and anaplerosis, the
CC processes that control the levels of metabolic intermediates in the
CC citric acid cycle (By similarity). At low glucose levels, it catalyzes
CC the cataplerotic conversion of oxaloacetate to phosphoenolpyruvate
CC (PEP), the rate-limiting step in the metabolic pathway that produces
CC glucose from lactate and other precursors derived from the citric acid
CC cycle (By similarity). At high glucose levels, it catalyzes the
CC anaplerotic conversion of phosphoenolpyruvate to oxaloacetate (By
CC similarity). Acts as a regulator of formation and maintenance of memory
CC CD8(+) T-cells: up-regulated in these cells, where it generates
CC phosphoenolpyruvate, via gluconeogenesis (By similarity). The resultant
CC phosphoenolpyruvate flows to glycogen and pentose phosphate pathway,
CC which is essential for memory CD8(+) T-cells homeostasis (By
CC similarity). In addition to the phosphoenolpyruvate carboxykinase
CC activity, also acts as a protein kinase when phosphorylated at Ser-90:
CC phosphorylation at Ser-90 by AKT1 reduces the binding affinity to
CC oxaloacetate and promotes an atypical serine protein kinase activity
CC using GTP as donor (By similarity). The protein kinase activity
CC regulates lipogenesis: upon phosphorylation at Ser-90, translocates to
CC the endoplasmic reticulum and catalyzes phosphorylation of INSIG
CC proteins (INSIG1 and INSIG2), thereby disrupting the interaction
CC between INSIG proteins and SCAP and promoting nuclear translocation of
CC SREBP proteins (SREBF1/SREBP1 or SREBF2/SREBP2) and subsequent
CC transcription of downstream lipogenesis-related genes (By similarity).
CC {ECO:0000250|UniProtKB:P35558, ECO:0000250|UniProtKB:Q9Z2V4,
CC ECO:0000269|PubMed:26792594}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=GTP + oxaloacetate = CO2 + GDP + phosphoenolpyruvate;
CC Xref=Rhea:RHEA:10388, ChEBI:CHEBI:16452, ChEBI:CHEBI:16526,
CC ChEBI:CHEBI:37565, ChEBI:CHEBI:58189, ChEBI:CHEBI:58702; EC=4.1.1.32;
CC Evidence={ECO:0000269|PubMed:26792594};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:10389;
CC Evidence={ECO:0000250|UniProtKB:Q9Z2V4};
CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:10390;
CC Evidence={ECO:0000250|UniProtKB:Q9Z2V4};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=GTP + L-seryl-[protein] = GDP + H(+) + O-phospho-L-seryl-
CC [protein]; Xref=Rhea:RHEA:64020, Rhea:RHEA-COMP:9863, Rhea:RHEA-
CC COMP:11604, ChEBI:CHEBI:15378, ChEBI:CHEBI:29999, ChEBI:CHEBI:37565,
CC ChEBI:CHEBI:58189, ChEBI:CHEBI:83421;
CC Evidence={ECO:0000250|UniProtKB:P35558};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:64021;
CC Evidence={ECO:0000250|UniProtKB:P35558};
CC -!- COFACTOR:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000250|UniProtKB:P35558};
CC Note=Binds 1 Mn(2+) ion per subunit. {ECO:0000250|UniProtKB:P35558};
CC -!- ACTIVITY REGULATION: Phosphoenolpyruvate carboxykinase activity is
CC regulated by acetylation and glucose levels (By similarity). The
CC anaplerotic conversion of phosphoenolpyruvate to oxaloacetate is
CC improved by PCK1 acetylation on Lys-91 (K91ac), Lys-473 (K473ac) and
CC Lys-521 (K521ac) (By similarity). High glucose concentrations favor
CC PCK1 anaplerotic activity by triggering acetylation on Lys-91 (K91ac).
CC At low glucose levels, SIRT1-mediated deacetylation of Lys-91 promotes
CC the cataplerotic conversion of oxaloacetate to phosphoenolpyruvate.
CC Phosphorylation at Ser-90 reduces the binding affinity to oxaloacetate
CC and converts the enzyme into an atypical protein kinase using GTP as
CC donor (By similarity). {ECO:0000250|UniProtKB:P07379,
CC ECO:0000250|UniProtKB:P35558, ECO:0000250|UniProtKB:Q9Z2V4}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=240 uM for phosphoenolpyruvate {ECO:0000269|PubMed:26792594};
CC KM=27.4 uM for GDP {ECO:0000269|PubMed:26792594};
CC KM=13.1 uM for oxaloacetate {ECO:0000269|PubMed:26792594};
CC KM=45.9 uM for GTP {ECO:0000269|PubMed:26792594};
CC Note=kcat is 6.8 sec(-1) with phosphoenolpyruvate as substrate
CC (PubMed:26792594). kcat is 7.6 sec(-1) with GDP as substrate
CC (PubMed:26792594). kcat is 46 sec(-1) with oxaloacetate as substrate.
CC kcat is 39 sec(-1) with GTP as substrate (PubMed:26792594).
CC {ECO:0000269|PubMed:26792594};
CC -!- PATHWAY: Carbohydrate biosynthesis; gluconeogenesis.
CC {ECO:0000250|UniProtKB:Q9Z2V4}.
CC -!- SUBUNIT: Monomer. {ECO:0000250|UniProtKB:P35558}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm, cytosol
CC {ECO:0000250|UniProtKB:P35558}. Endoplasmic reticulum
CC {ECO:0000250|UniProtKB:P35558}. Note=Phosphorylation at Ser-90 promotes
CC translocation to the endoplasmic reticulum.
CC {ECO:0000250|UniProtKB:P35558}.
CC -!- DOMAIN: Binds oxysterols in a pocket within their transmembrane domains
CC and interacts with SCAP via transmembrane domains 3 and 4.
CC {ECO:0000250|UniProtKB:Q9Y5U4}.
CC -!- DOMAIN: The KxHxx motif mediates association with the coatomer complex.
CC {ECO:0000250|UniProtKB:Q9Y5U4}.
CC -!- PTM: Acetylated (By similarity). Lysine acetylation by p300/EP300 is
CC increased on high glucose conditions and promotes ubiquitination by
CC UBR5, acetylation is enhanced in the presence of BAG6. Deacetylated by
CC SIRT2 (By similarity). Deacetylated by SIRT1 (By similarity).
CC {ECO:0000250|UniProtKB:P35558, ECO:0000250|UniProtKB:Q9Z2V4}.
CC -!- PTM: Ubiquitination by UBR5 leads to proteasomal degradation.
CC {ECO:0000250|UniProtKB:P35558}.
CC -!- PTM: Phosphorylated in a GSK3B-mediated pathway; phosphorylation
CC affects the efficiency of SIRT1-mediated deacetylation, and regulates
CC PCK1 ubiquitination and degradation. Phosphorylation at Ser-90 by AKT1
CC reduces the binding affinity to oxaloacetate and promotes the protein
CC kinase activity: phosphorylated PCK1 translocates to the endoplasmic
CC reticulum, where it phosphorylates INSIG1 and INSIG2.
CC {ECO:0000250|UniProtKB:P35558}.
CC -!- MISCELLANEOUS: In eukaryotes there are two isozymes: a cytoplasmic one
CC and a mitochondrial one. {ECO:0000250|UniProtKB:Q9Z2V4}.
CC -!- SIMILARITY: Belongs to the phosphoenolpyruvate carboxykinase [GTP]
CC family. {ECO:0000305}.
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DR EMBL; AEMK02000106; -; NOT_ANNOTATED_CDS; Genomic_DNA.
DR EMBL; FJ668384; ACR66221.1; -; Genomic_DNA.
DR EMBL; CR956387; CAN13128.1; -; Genomic_DNA.
DR RefSeq; NP_001116630.1; NM_001123158.1.
DR RefSeq; XP_005673100.1; XM_005673043.2.
DR AlphaFoldDB; A0A4X1UM84; -.
DR SMR; A0A4X1UM84; -.
DR STRING; 9823.ENSSSCP00000008003; -.
DR Ensembl; ENSSSCT00025102441; ENSSSCP00025045327; ENSSSCG00025074360.
DR GeneID; 100144531; -.
DR KEGG; ssc:100144531; -.
DR CTD; 5105; -.
DR eggNOG; KOG3749; Eukaryota.
DR HOGENOM; CLU_028872_1_1_1; -.
DR OMA; XNLRAIN; -.
DR OrthoDB; 286671at2759; -.
DR TreeFam; TF314402; -.
DR Reactome; R-SSC-70263; Gluconeogenesis.
DR UniPathway; UPA00138; -.
DR Proteomes; UP000008227; Unplaced.
DR Proteomes; UP000314985; Unplaced.
DR GO; GO:0005829; C:cytosol; ISS:UniProtKB.
DR GO; GO:0005783; C:endoplasmic reticulum; ISS:UniProtKB.
DR GO; GO:0031406; F:carboxylic acid binding; IEA:Ensembl.
DR GO; GO:0005525; F:GTP binding; IEA:UniProtKB-KW.
DR GO; GO:0000287; F:magnesium ion binding; IEA:Ensembl.
DR GO; GO:0030145; F:manganese ion binding; IEA:Ensembl.
DR GO; GO:0004613; F:phosphoenolpyruvate carboxykinase (GTP) activity; IDA:UniProtKB.
DR GO; GO:0106264; F:protein serine kinase activity (using GTP as donor); ISS:UniProtKB.
DR GO; GO:0071333; P:cellular response to glucose stimulus; IEA:Ensembl.
DR GO; GO:0032869; P:cellular response to insulin stimulus; ISS:UniProtKB.
DR GO; GO:0051365; P:cellular response to potassium ion starvation; IEA:Ensembl.
DR GO; GO:0006094; P:gluconeogenesis; IEA:UniProtKB-UniPathway.
DR GO; GO:0046327; P:glycerol biosynthetic process from pyruvate; IEA:Ensembl.
DR GO; GO:0006107; P:oxaloacetate metabolic process; IDA:UniProtKB.
DR GO; GO:0018105; P:peptidyl-serine phosphorylation; ISS:UniProtKB.
DR GO; GO:0043382; P:positive regulation of memory T cell differentiation; ISS:UniProtKB.
DR GO; GO:0061402; P:positive regulation of transcription from RNA polymerase II promoter in response to acidic pH; IEA:Ensembl.
DR GO; GO:0046890; P:regulation of lipid biosynthetic process; ISS:UniProtKB.
DR GO; GO:0009617; P:response to bacterium; IEA:Ensembl.
DR CDD; cd00819; PEPCK_GTP; 1.
DR Gene3D; 3.40.449.10; -; 1.
DR Gene3D; 3.90.228.20; -; 1.
DR HAMAP; MF_00452; PEPCK_GTP; 1.
DR InterPro; IPR018091; PEP_carboxykin_GTP_CS.
DR InterPro; IPR013035; PEP_carboxykinase_C.
DR InterPro; IPR008209; PEP_carboxykinase_GTP.
DR InterPro; IPR035077; PEP_carboxykinase_GTP_C.
DR InterPro; IPR035078; PEP_carboxykinase_GTP_N.
DR InterPro; IPR008210; PEP_carboxykinase_N.
DR PANTHER; PTHR11561; PTHR11561; 1.
DR Pfam; PF00821; PEPCK_GTP; 1.
DR Pfam; PF17297; PEPCK_N; 1.
DR PIRSF; PIRSF001348; PEP_carboxykinase_GTP; 1.
DR SUPFAM; SSF68923; SSF68923; 1.
DR PROSITE; PS00505; PEPCK_GTP; 1.
PE 1: Evidence at protein level;
KW Acetylation; Cytoplasm; Decarboxylase; Endoplasmic reticulum;
KW Gluconeogenesis; GTP-binding; Kinase; Lyase; Manganese; Metal-binding;
KW Nucleotide-binding; Phosphoprotein; Reference proteome; Transferase;
KW Ubl conjugation.
FT CHAIN 1..622
FT /note="Phosphoenolpyruvate carboxykinase, cytosolic [GTP]"
FT /id="PRO_0000450692"
FT REGION 457..487
FT /note="Omega-loop"
FT /evidence="ECO:0000250|UniProtKB:P07379"
FT ACT_SITE 288
FT /evidence="ECO:0000250|UniProtKB:P35558"
FT BINDING 87
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P35558"
FT BINDING 235..237
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P35558"
FT BINDING 244
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000250|UniProtKB:P35558"
FT BINDING 264
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000250|UniProtKB:P35558"
FT BINDING 286
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P07379"
FT BINDING 287..292
FT /ligand="GTP"
FT /ligand_id="ChEBI:CHEBI:37565"
FT /evidence="ECO:0000250|UniProtKB:P35558"
FT BINDING 311
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000250|UniProtKB:P35558"
FT BINDING 403..405
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P35558"
FT BINDING 405
FT /ligand="GTP"
FT /ligand_id="ChEBI:CHEBI:37565"
FT /evidence="ECO:0000250|UniProtKB:P35558"
FT BINDING 436
FT /ligand="GTP"
FT /ligand_id="ChEBI:CHEBI:37565"
FT /evidence="ECO:0000250|UniProtKB:P35558"
FT BINDING 530..533
FT /ligand="GTP"
FT /ligand_id="ChEBI:CHEBI:37565"
FT /evidence="ECO:0000250|UniProtKB:P35558"
FT MOD_RES 70
FT /note="N6-acetyllysine"
FT /evidence="ECO:0000250|UniProtKB:P35558"
FT MOD_RES 71
FT /note="N6-acetyllysine"
FT /evidence="ECO:0000250|UniProtKB:P35558"
FT MOD_RES 90
FT /note="Phosphoserine"
FT /evidence="ECO:0000250|UniProtKB:P35558"
FT MOD_RES 91
FT /note="N6-acetyllysine"
FT /evidence="ECO:0000250|UniProtKB:P35558"
FT MOD_RES 118
FT /note="Phosphoserine"
FT /evidence="ECO:0000250|UniProtKB:Q9Z2V4"
FT MOD_RES 286
FT /note="Phosphoserine"
FT /evidence="ECO:0000250|UniProtKB:Q16822"
FT MOD_RES 473
FT /note="N6-acetyllysine"
FT /evidence="ECO:0000250|UniProtKB:P07379"
FT MOD_RES 521
FT /note="N6-acetyllysine"
FT /evidence="ECO:0000250|UniProtKB:P07379"
FT MOD_RES 594
FT /note="N6-acetyllysine"
FT /evidence="ECO:0000250|UniProtKB:P35558"
FT VARIANT 139
FT /note="M -> L (impaired phosphoenolpyruvate carboxykinase
FT activity, leading to reduced intramuscular fat content,
FT enhanced backfat thickness and lower meat quality)"
FT /evidence="ECO:0000269|PubMed:26792594"
SQ SEQUENCE 622 AA; 68884 MW; 277ECBFCEF862B66 CRC64;
MPPQLSNGLN HSAKVVRGTL DSLPQAVRDF VESSAKLCQP DQIHICDGSE EENQQLLSHM
EEEGVIKRLK KYDNCWLALT DPRDVARIES KTVIITQEQR DAVPIPRSGL SQLGRWMSPE
DFEKAFNARF PGCMKGRTMY VIPFSMGPLG SPLSKIGIEL TDSPYVVASM RIMTRMGSAV
LDALGAGEFI KGLHSVGCPL PLKKPLVNNW ACNPELTLIA HLPDRREIIS FGSGYGGNSL
LGKKCFALRM ASRLAKEEGW LAEHMLILGV TNPEGQKKYF AAAFPSACGK TNLAMMNPTL
PGWKVECVGD DIAWMKFDQQ GNLRAINPEN GFFGVAPGTS VKTNPNAIKT IQSNTIFTNV
AETSDGGVYW EGIDQPLAPG IKLTSWKGTE WDPKDGEPCA HPNSRFCTPA SQCPIIDPAW
EAPEGVPIEG IIFGGRRPAG VPLVYEALSW QHGVFVGAAM RSEATAAAEH KGKVIMHDPF
AMRPFFGYNF GKYLAHWLSM AQHPAAKLPK IFHVNWFRKD KDGRFLWPGF GENCRVLAWM
FDRVQGKGGA RLTPIGYVPE EAALDLRGLE AIRVSELFQV SKEFWEEEAD EIHKYLEEQV
NADLPYEIQS EVLALKQRIS QM
