Product Details

Purity

Greater than 90% as determined by SDS-PAGE.

Target Names

PCK1

Uniprot No.

P35558

Research Area

Metabolism

Alternative Names

cytosolic [GTP]; GTP; PCK1; PCKGC_HUMAN; PEP carboxykinase; PEPCK-C; PEPCK1; PEPCKC; Phosphoenolpyruvate carboxykinase 1 (soluble); Phosphoenolpyruvate carboxykinase 1; Phosphoenolpyruvate carboxykinase; Phosphoenolpyruvate carboxykinase, cytosolic [GTP]; Phosphoenolpyruvate carboxykinase, cytosolic; Phosphoenolpyruvate carboxylase; Phosphopyruvate carboxylase

Species

Homo sapiens (Human)

Source

E.coli

Expression Region

1-622aa

Target Protein Sequence

MPPQLQNGLNLSAKVVQGSLDSLPQAVREFLENNAELCQPDHIHICDGSEEENGRLLGQMEEEGILRRLKKYDNCWLALTDPRDVARIESKTVIVTQEQRDTVPIPKTGLSQLGRWMSEEDFEKAFNARFPGCMKGRTMYVIPFSMGPLGSPLSKIGIELTDSPYVVASMRIMTRMGTPVLEAVGDGEFVKCLHSVGCPLPLQKPLVNNWPCNPELTLIAHLPDRREIISFGSGYGGNSLLGKKCFALRMASRLAKEEGWLAEHMLILGITNPEGEKKYLAAAFPSACGKTNLAMMNPSLPGWKVECVGDDIAWMKFDAQGHLRAINPENGFFGVAPGTSVKTNPNAIKTIQKNTIFTNVAETSDGGVYWEGIDEPLASGVTITSWKNKEWSSEDGEPCAHPNSRFCTPASQCPIIDAAWESPEGVPIEGIIFGGRRPAGVPLVYEALSWQHGVFVGAAMRSEATAAAEHKGKIIMHDPFAMRPFFGYNFGKYLAHWLSMAQHPAAKLPKIFHVNWFRKDKEGKFLWPGFGENSRVLEWMFNRIDGKASTKLTPIGYIPKEDALNLKGLGHINMMELFSISKEFWEKEVEDIEKYLEDQVNADLPCEIEREILALKQRISQM
Note: The complete sequence including tag sequence, target protein sequence and linker sequence could be provided upon request.

Mol. Weight

73.2kDa

Protein Length

Full Length

Tag Info

N-terminal 6xHis-tagged

Form

Liquid or Lyophilized powder
Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand.

Buffer

If the delivery form is liquid, the default storage buffer is Tris/PBS-based buffer, 5%-50% glycerol.
Note: If you have any special requirement for the glycerol content, please remark when you place the order.
If the delivery form is lyophilized powder, the buffer before lyophilization is Tris/PBS-based buffer, 6% Trehalose, pH 8.0.

Reconstitution

We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final concentration of glycerol is 50%. Customers could use it as reference.

Troubleshooting and FAQs

Protein FAQs

Storage Condition

Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.

Shelf Life

The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.

Lead Time

3-7 business days

Notes

Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.

Datasheet & COA

Please contact us to get it.

Description

This Human PCK1 recombinant protein was produced in E.coli, where the gene sequence encoding Human PCK1 (1-622aa) was expressed with the N-terminal 6xHis tag. The purity of this PCK1 protein was greater than 90% by SDS-PAGE.
PCK1 plays a critical role in the gluconeogenesis pathway. This pathway helps maintain blood sugar levels when they are low due to fasting or extended periods without eating. It allows liver and kidney cells to synthesize glucose from non-sugar substrates to supply to other tissues, including the brain. The activity of PCK1 is regulated by various factors, including hormones such as insulin and adrenaline, energy status, and glucose concentration. Insulin inhibits the activity of PCK1, promoting glucose uptake and storage, while stress hormones like adrenaline can stimulate PCK1 activity, increasing glucose release. PCK1 is crucial for maintaining normal blood sugar levels and energy balance. Defects or abnormal activity of PCK1 are associated with diabetes, obesity, and metabolic disorders. Due to its importance in glucose metabolism, PCK1 has become a potential drug target. Some research is exploring the possibility of regulating PCK1 activity to treat diseases related to blood sugar regulation.

Customer Reviews and Q&A

■ Customer Reviews

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■ Q&A

I wanted to mention a critical thing about this protein. It needs Manganese (Mn2+) to fold properly. We found that our in house protein was behaving strangely in the presence of inhibitors and this was due to the presence of aberrantly folded protein. We had prepared His tagged material and to purify it metal affinity chromatography was used which depletes the protein from any Mn2+. I would highly recommend adding MnCl2 (1-2mM) to the final product preferably immediately after purification. Particularly if you are using metal affinity resin (talon or some other). Even if you are not I would strongly suggest inclusion of Manganese in the final buffer prior to freezing and shipment.
And:
One other point. The enzyme has a very active cysteine residue in the active site that is required for catalysis. It needs to be reduced. So use DDT (2-5 mM) or TCEP (1mM) in the final buffer.
Please let me know if adding MnCl2 and DDT or TCEP to the final product will be possible.

Thank you for notifying us of these key information in time. We will add MnCl2 and DDT or TCEP to the final product.

Target Background

Function

Cytosolic phosphoenolpyruvate carboxykinase that catalyzes the reversible decarboxylation and phosphorylation of oxaloacetate (OAA) and acts as the rate-limiting enzyme in gluconeogenesis. Regulates cataplerosis and anaplerosis, the processes that control the levels of metabolic intermediates in the citric acid cycle. At low glucose levels, it catalyzes the cataplerotic conversion of oxaloacetate to phosphoenolpyruvate (PEP), the rate-limiting step in the metabolic pathway that produces glucose from lactate and other precursors derived from the citric acid cycle. At high glucose levels, it catalyzes the anaplerotic conversion of phosphoenolpyruvate to oxaloacetate. Acts as a regulator of formation and maintenance of memory CD8(+) T-cells: up-regulated in these cells, where it generates phosphoenolpyruvate, via gluconeogenesis. The resultant phosphoenolpyruvate flows to glycogen and pentose phosphate pathway, which is essential for memory CD8(+) T-cells homeostasis. In addition to the phosphoenolpyruvate carboxykinase activity, also acts as a protein kinase when phosphorylated at Ser-90: phosphorylation at Ser-90 by AKT1 reduces the binding affinity to oxaloacetate and promotes an atypical serine protein kinase activity using GTP as donor. The protein kinase activity regulates lipogenesis: upon phosphorylation at Ser-90, translocates to the endoplasmic reticulum and catalyzes phosphorylation of INSIG proteins (INSIG1 and INSIG2), thereby disrupting the interaction between INSIG proteins and SCAP and promoting nuclear translocation of SREBP proteins (SREBF1/SREBP1 or SREBF2/SREBP2) and subsequent transcription of downstream lipogenesis-related genes.

Gene References into Functions

This study reveals a unique mechanism to suppress hepatocellular carcinoma by switching from glycolysis to gluconeogenesis through Nur77 antagonism of PEPCK1 degradation. PMID: 28240261
circC3P1 acts as a tumor suppressor via enhancing PCK1 expression by sponging miR-4641 in hepatocellular carcinoma. PMID: 29608893
The expression levels of PCK1 were suppressed in hepatocellular carcinoma (HCC) samples and in cells derived from HCC tissues. PMID: 29768256
Study investigated the molecular basis of such effects focusing on a commonly studied polymorphism in pig Pgc1alpha, which changes a cysteine at position 430 (WT) of the protein to a serine (C430S); found that differential O-GlcNAcylation of Pgc1alpha regulates PCK1 activity and this molecular mechanism could explain at least in part the epistatic interaction between both genes. PMID: 28644880
Study identified a novel homozygous PCK1 missense mutation causing cytosolic phosphoenolpyruvate carboxykinase deficiency presenting as childhood hypoglycemia. PMID: 28216384
HBXIP is able to depress the gluconeogenesis in hepatoma cells by suppressing PCK1 to promote hepatocarcinogenesis, involving miR-135a/FOXO1 axis and PI3K/Akt/p-FOXO1 pathway. PMID: 27609066
an extended binding site in the catalytic cleft of cPEPCK which is used by 3-MPA to inhibit cPEPCK non-competitively. PMID: 26528723
A mutation of PCK1 associated with Smith-Magenis Syndrome, cytosolic PEPCK deficiency and NMDA receptor glutamate insensitivity. PMID: 24863970
Insulin-dependent translocation of FOXO1 regulated transcriptional repression of PCK1 concomitant with the formation of the FOXO1-euchromatic histone-lysine N-methyltransferase2 (EHMT2) complex and histone modifications of the PCK1 promoter region PMID: 25736587
Lipopolysaccharide and monophosphoryl lipid A also up-regulated G6PC and PCK1 transcript abundance in a TLR4-dependent manner. PMID: 23465595
The PCK1 tag-single nucleotide polymorphisms influenced insulin resistance by interacting with plasma n-3 polyunsaturated fatty acids levels in metabolic syndrome patients. PMID: 23092637
HCV core protein expression-mediated FOXO1 activation and the increased PGC-1alpha leaded to the elevation of PCK1 at the mRNA level PMID: 22489460
Acetylation regulates the stability of the gluconeogenic rate-limiting enzyme PEPCK1. PMID: 21726808
Data show PEPCK-C and CPT-1 mRNAs are more abundant in non-tumoral tissues than in the tumoral counterpart, whereas the opposite occurred for the FAS gene. PMID: 20691246
Studies demonstrate the association between PCK1 and smaller average brain volume. PMID: 21152065
Study in the EYHS cohort failed to identify an association of PCK1 polymorphisms with obesity, physical activity, and fitness. PMID: 20134411
crystal structure of human cytosolic phosphoenolpyruvate carboxykinase reveals a new GTP-binding site PMID: 11851336
Conserved amino acids within CCAAT enhancer-binding proteins (C/EBP(alpha) and beta) regulate gene expression PMID: 11997389
promoter context may influence which domains within a transcription factor are employed to mediate transactivation of PEPCK PMID: 12237288
the PGC- 1-regulated phospho(enol)pyruvate carboxykinase (PEPCK) promoter is a novel target promoter for SHP inhibition PMID: 12324453
transcription of the gene for phosphoenolpyruvate carboxykinase (GTP) in the liver is regulated by SREBP-1c and Sp1 PMID: 14744869
Promoter single nucleotide polymorphism in PCK1 was resistant to down-regulation by insulin in vitro and associated with type 2 diabetes mellitus. PMID: 14764811
PCK1 has been implicated as one of many genes associated with type 2 diabetes mellitus [review] PMID: 15046742
AUF1 binds to multiple destabilizing elements within the 3'-UTR that participate in the rapid turnover of the phosphoenolpyruvate carboxykinase mRNA. PMID: 15951444
The multiple instability elements present within the 3'-UTR may function synergistically to mediate both the rapid degradation and the cAMP-induced stabilization of PEPCK mRNA. The latter process may result from a PKA-dependent phosphorylation of AUF1. PMID: 16144962
Subjects with the PCK1-232G/G genotype had more carotid IMT (0.80+/-0.02 versus 0.73+/-0.03 mm; P=0.007) but less TPV (0.10+/-0.09 versus 0.38+/-0.13; P=0.03) than subjects with other genotypes. PMID: 16282543
Significant association of single nucleotide polymorphism within three genes--PPARgamma, SOS2, and PCK1--with Alzheimer's disease, was confirmed. PMID: 17440948
IFN-gamma exerts a tissue-specific action in rodents and humans, having glyceroneogenesis and the PEPCK-C gene as selective targets to intensify fatty acids release from adipocytes PMID: 17495004
The effects of several variants of retinoic acid on the expression and activity of PEPCK in human, mouse, and rat adipose tissue are reported. PMID: 18492826
Reduced CREB phosphorylation (Ser-129) associated with inactivation of GSK3beta by Ser-9 phosphorylation may be the major mechanism underlying PEPCK-C gene suppression by AMPK-activating agents such as biguanide PMID: 18801732
The aromatic ring of Tyr235 in PEPCK helps to position PEP in the active site and the hydroxyl group allows an optimal PEP-Mn(2+) distance for efficient phosphoryl transfer and overall catalysis. PMID: 19021757
Val184 of PCK1 gene might increase the risk of type 2 diabetes in eastern Chinese population with BMI<23 kg/m(2) via reducing the PEPCK-C activity. PMID: 19070910
Study validated a predicted SP1 binding site in the control of PCK1 transcription using gel shift and reporter assays. PMID: 19329064
study is the first to investigate the association between variants of the PCK1 gene and type 2 diabetes in South Asians PMID: 19725958

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Involvement in disease

Phosphoenolpyruvate carboxykinase deficiency, cytosolic (PCKDC)

Subcellular Location

Cytoplasm, cytosol. Endoplasmic reticulum.

Protein Families

Phosphoenolpyruvate carboxykinase [GTP] family

Tissue Specificity

Major sites of expression are liver, kidney and adipocytes.

Database Links

HGNC: 8724

OMIM: 261680

KEGG: hsa:5105

STRING: 9606.ENSP00000319814

UniGene: Hs.1872

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