Recombinant Human PGM1 His-tag Protein, CF

R&D Systems | Catalog # 11681-P1

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human PGM1 His-tag Protein (11681-P1)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

E. coli

Accession Number

P36871.3

Applications

Enzyme Activity
View all PGM1 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human PGM1 protein
Val2-Thr562, with an N-terminal Met and 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

62 kDa

SDS-PAGE

58-64 kDa, under reducing conditions

Activity

Measured by its ability to convert alpha -D-glucose 1-phosphate to alpha -D-glucose 6-phosphate in a coupled reaction.  
The specific activity is >6500 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Human PGM1 His-tag Protein, CF

Recombinant Human PGM1 His-tag Protein SDS-PAGE.

2 μg/lane of Recombinant Human PGM1 His-tag Protein (Catalog # 11681-P1) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 58-64 kDa, under reducing conditions.

Formulation, Preparation, and Storage

11681-P1
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and TCEP.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: PGM1

Phosphoglucomutase-1 (PGM1) has a central role in metabolism as it mediates the switch between glycolysis and gluconeogenesis by catalyzing the interconversion of glucose 1-phosphate and glucose 6-phosphate within the nucleotide sugar, galactose, and pentose phosphate pathways and is involved in protein N-glycosylation (1). It is a member of the phosphohexose mutase family and is ubiquitously expressed in tissues and organs (1,2). PGM1 is a cytosolic, magnesium-dependent monomer with four domains that form an overall heart shape (1,2). The active site is located in a large central cleft in the middle of the four domains that contain a catalytic phosphoserine that participates in phosphoryl transfer, a metal binding loop, a sugar-binding loop, and a phosphate-binding site (2).  Deficiency in PGM1 activity has been identified to cause glycogen storage disorder GSD XIV and is also a rare genetic disorder congenital disorder of glycosylation (CDG) called PGM1-CDG or CDG1T (3-6) with affected individuals showing multiple disease phenotypes including cardiomyopathy, muscular weakness, and hepatopathy (4,6,7). Some, but not all, clinical symptoms can be improved with oral D-galactose treatment (4,7,8). In addition, PGM1 is down-regulated in cancers such as hepatocellular carcinoma (HCC) and colorectal cancer (CRC) and up-regulated cancers such as lung and gastric cancer where it has been linked to lower survival in all cases making PGM1 a potential biomarker in several cancers (9-12). Galactose supplementation was not corrective in HCC suggesting PGM1's role in regulation of glycogen synthesis and glucose regulation is critical in promotion of tumor cell proliferation and growth (9,10,12,13). Given the key role PGM1 plays in metabolism in cancer and disease there is interest in PGM1 as a therapeutic target (10-12). 

References

  1. Stiers, K.M. et al. (2016) J. Mol. Biol. 428:1493. 
  2. Shackelford, G.S. et al. (2004) Protein Sci. 13:2130.
  3. Lee, Y. et al. (2014) J. Biol. Chem. 289:32010.
  4. Tegtmeyer, L.C. et al. (2014) N. Engl. J. Med. 370:533.
  5. Beamer, L.J. (2015) J. Inherit Metab. Dis. 38:243.
  6. Altassan, R et al. (2021) J. Inherit. Metab. Dis. 44:148.
  7. Fernlund, E. et al. (2019) Forensic Sci. Int. Genet. 43:102111.
  8. Morava, E. (2014) Mol. Genet. Metab. 112:275.
  9. Jin, G.Z. et al. (2018) PloS Biol. 16:e2006483.
  10. Li, Y. et al. (2020) Cancer Lett. 478:82.
  11. Cao, B. et al. (2021) Cancer Cell Int. 21:481.
  12. Zheng, Z. et al. (2022) Cancer Cell Int. 22:201. 
  13. Zheng, Z. et al. (2022) Front. Oncol. 12:1060372. 

Long Name

phosphoglucomutase 1

Alternate Names

CDG1T, GSD14

Entrez Gene IDs

5236 (Human); 72157 (Mouse); 24645 (Rat)

Gene Symbol

PGM1

UniProt

P36871.3

Additional PGM1 Products

Product Documents for Recombinant Human PGM1 His-tag Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Human PGM1 His-tag Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Human PGM1 His-tag Protein, CF (11681-P1):

Materials
  • Assay Buffer: 25 mM Tris, 1 mM MgCl2, pH 8.0
  • Recombinant Human PGM1 His-tag (rhPGM1) (Catalog # 11681-PI)
  • Coupling Enzyme: Recombinant Human G6PD His-tag (rhG6PD)  (Catalog # 10096-DH)
  • L-Cysteine, 50 mM stock in deionized water
  • beta -Nicotinamide adenine dinucleotide phosphate (NADP+), 50 mM stock in deionized water
  • Substrate: alpha -D-Glucose 1-phosphate, 10 mM stock in deionized water
  • Clear 96-well Plate
  • Plate Reader with Absorbance Read Capability
  1. Create a Master Mix containing 2 mM L-Cysteine, 2 mM NADP+, 5 µg/mL rhG6PD, and 0.1 µg/mL rhPGM1 in Assay Buffer. Include a Control containing the same master mix components in Assay Buffer, but without rhPGM1.
  2. Incubate the Master Mixes and Control at room temperature for 10 minutes.
  3. Dilute Substrate to 1 mM in Assay Buffer.
  4. Following incubation, load 50 µL of each master mix (including the Control) to the plate and start the reaction by adding 50 µL of 1 mM Substrate to all wells. 
  5. Read plate at 340 nm (absorbance) in kinetic mode for 5 minutes.
  6. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol
ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)

     

*Adjusted for Control
**Using the extinction coefficient 6220 M-1cm-1
***Using the path correction 0.32 cm
Note: The output of many spectrophotometers is in mOD.

Per Well:

  • rhPGM1: 0.005 µg
  • rhG6PD: 0.250 µg
  • Substrate: 0.5 mM
  • L-Cysteine: 1 mM
  • NADP+: 1 mM












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