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Literature summary for 2.7.1.40 extracted from

  • Kumar, S.; Patel, A.K.
    Purification and characterization of prolyl hydroxylase 3/pyruvate kinase isoform 2 protein complex (2020), Mol. Biotechnol., 62, 111-118 .
    View publication on PubMed
Search for more literature for 2.7.1.40

Cloned(Commentary)

Cloned (Comment) Organism
gene PKM2, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)pLys Homo sapiens

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant PHD3/PKM2 complex, hanging drop vapor diffusion method, mixing of 5 mg/ml protein in 20 mM MES, pH 6.0, 75 mM NaCl, and 1 mM DTT, in a 1:1 ration with reservoir solution, 18°C, two weeks, X-ray diffraction structure determination and analysis at 3.0 A resolution, molecular replacement method and modeling Homo sapiens

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
60000
-
 recombinant enzyme, gel filtration Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ADP + phosphoenolpyruvate Homo sapiens
-
 ATP + pyruvate
-
 ?
additional information Homo sapiens direct physical binding through protein-protein interaction between PHD3 and PKM2 is observed. Homology three-dimensional model of PHD3/PKM2 complex ?
-
-

Organism

Organism UniProt Comment Textmining
Homo sapiens P14618
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3)pLys by nickel affinity chromatography, the His6-tag is cleaved off by PreScission protease followed by a second step of nickel affinity chromatography, ultrafiltration, and gel filtration Homo sapiens

Source Tissue

Source Tissue Comment Organism Textmining
carcinoma cell
-
 Homo sapiens
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ADP + phosphoenolpyruvate
-
 Homo sapiens ATP + pyruvate
-
 ?
additional information direct physical binding through protein-protein interaction between PHD3 and PKM2 is observed. Homology three-dimensional model of PHD3/PKM2 complex Homo sapiens ?
-
-

Subunits

Subunits Comment Organism
? x * 58000, recombinant detagged enzyme, SDS-PAGE Homo sapiens

Synonyms

Synonyms Comment Organism
PKM2
-
 Homo sapiens
pyruvate kinase isoforms 2
-
 Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Homo sapiens

General Information

General Information Comment Organism
physiological function The prolyl hydroxylase 3 (PHD3, EC 1.14.11.29) protein is less abundant in normal oxygen conditions (normoxia) but increases under deficient oxygen condition (hypoxia). Since cancerous cells often thrive in hypoxic conditions and predominantly express the pyruvate kinase isoforms 2 (PKM2), the PHD3/PKM2 interaction might be particularly important in cancer development. Protein interaction analysis and PHD3/PKM2 complex structure analysis, overview. PHD3 hydroxylates the PKM2 at two specific proline residues. The hydroxylated PKM2 shows enhanced binding with HIF-1alpha, which in turn increases the activity of HIF-1alpha Homo sapiens