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Literature summary extracted from

  • Wang, X.; Engel, P.
    An optimised system for refolding of human glucose 6-phosphate dehydrogenase (2009), BMC Biotechnol., 9, 19.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.1.1.49 6-aminohexanoic acid marginally increases the refolding yield in the presence of Arg, and it is unable to replace Arg in promoting a high refolding yield Homo sapiens
1.1.1.49 butanol marginally increases the refolding yield in the presence of Arg, and it is unable to replace Arg in promoting a high refolding yield Homo sapiens
1.1.1.49 cyclophilin A facilitates refolding, the yield, at about 82%, is about 12% higher than without cyclophilin A. Even with 0.0004 mM cyclophilin A in the standard refolding buffer, maximal recovery takes about 7 days Homo sapiens
1.1.1.49 dithiothreitol dithiothreitol has major positive influence on refolding Homo sapiens
1.1.1.49 glycerol marginally increases the refolding yield in the presence of Arg, and it is unable to replace Arg in promoting a high refolding yield Homo sapiens
1.1.1.49 L-Arg L-Arg has major positive influence on refolding. L-Arg is the key player in the refolding of human G6PD, preventing the aggregation of folding intermediate Homo sapiens
1.1.1.49 additional information temperature and protein concentration have positive influence on refolding. Refolding at 25°C is relatively slow, taking about one week to reach the maximum activity. Final refolding yield at pH 7.5 is about 69%, similar to 71% at pH 8.0, and only slightly higher than 65% at pH 8.5. At 30°C, refolding is faster in the early stages than at 25°C, but the highest activities, achieved after 4-5 days, are 49% at pH 7.5, 53% at pH 8.0, and 48% at pH 8.5, respectively, considerably lower than at 25°C. Correct refolding is severely suppressed at 37°C, with the highest recovery yield at 13%. Recovery yields are 54%, 70%, 56%, and 33% for protein concentrations at 5 microg/ml, 10 microg/ml, 20 microg/ml and 50 microg/ml, respectively Homo sapiens
1.1.1.49 PEG 3350 marginally increases the refolding yield in the presence of Arg, and it is unable to replace Arg in promoting a high refolding yield Homo sapiens
1.1.1.49 trehalose marginally increases the refolding yield in the presence of Arg, and it is unable to replace Arg in promoting a high refolding yield Homo sapiens

Application

EC Number Application Comment Organism
1.1.1.49 medicine refolding protocol can be applied to produce high recovery yield of folded protein with unaltered properties, paving the way for future studies on clinical G6PD mutants with folding defects and providing a useful model system to study the folding process of oligomeric proteins Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.49 guanidinium hydrochloride in the presence of 4 M, after 2 hours all secondary structure is lost Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.49 0.00694
-
 NADP+ refolded G6PD Homo sapiens
1.1.1.49 0.00707
-
 NADP+ native G6PD Homo sapiens
1.1.1.49 0.052
-
 D-glucose 6-phosphate native G6PD Homo sapiens
1.1.1.49 0.0547
-
 D-glucose 6-phosphate refolded G6PD Homo sapiens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.49 Na2HPO4 at 400 mM regains 22% activity on refolding Homo sapiens
1.1.1.49 Na2SO4 at 400 mM regains 23% activity on refolding Homo sapiens
1.1.1.49 NaCl at 400 mM regains 63% activity on refolding Homo sapiens
1.1.1.49 NaClO4 at 400 mM regains 2.3% activity on refolding Homo sapiens
1.1.1.49 NaSCN at 400 mM regains 1.4% activity on refolding Homo sapiens
1.1.1.49 Sodium acetate at 400 mM regains 58% activity on refolding Homo sapiens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.49 100000
-
 gel filtration, refolded protein Homo sapiens
1.1.1.49 200000
-
 gel filtration, native enzyme Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.49 Homo sapiens
-
-
-

Source Tissue

EC Number Source Tissue Comment Organism Textmining

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.49 D-glucose 6-phosphate + NADP+
-
 Homo sapiens D-glucono-1,5-lactone 6-phosphate + NADPH + H+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.49 dimer gel filtration, the refolded protein intermediates shift from dominant monomer to dimer, the gradual emergence of dimer correlating well with the regain of enzyme activity Homo sapiens
1.1.1.49 tetramer gel filtration Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
1.1.1.49 G6PD
-
 Homo sapiens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.49 275
-
 D-glucose 6-phosphate native G6PD Homo sapiens
1.1.1.49 279
-
 D-glucose 6-phosphate refolded G6PD Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.49 NADP+ NADP+ has major positive influence on refolding. NADP+ is essential for the folding intermediate to adopt native structure. Without added NADP+, the recovery yield is only about 10%. As the concentration of NADP+ increases, more enzyme activity can be regained in a concentration dependent manner Homo sapiens