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Literature summary extracted from

  • Sparla, F.; Fermani, S.; Falini, G.; Zaffagnini, M.; Ripamonti, A.; Sabatino, P.; Pupillo, P.; Trost, P.
    Coenzyme site-directed mutants of photosynthetic A4-GAPDH show selectively reduced NADPH-dependent catalysis, similar to regulatory AB-GAPDH inhibited by oxidized thioredoxin (2004), J. Mol. Biol., 340, 1025-1037.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.2.1.13 expression in Escherichia coli BL21 Spinacia oleracea

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.2.1.13 hanging-drop vapour-diffusion method is used to grow crystals of recombinant A4-GAPDH, T33A and S188A A4-GAPDH mutants complexed with NADP+ Spinacia oleracea

Protein Variants

EC Number Protein Variants Comment Organism
1.2.1.13 S188A affinity for NADPH is significantly decreased, decrease in the ratio of turnover number to Km-value in the NADPH-dependent reaction, significant expansion of the A4-tetramer Spinacia oleracea
1.2.1.13 T33A affinity for NADPH is significantly decreased, turnover-number for NADPH is lowered Spinacia oleracea
1.2.1.13 T33A/S188A affinity for NADPH is significantly decreased, turnover-number for NADPH is lowered Spinacia oleracea

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2.1.13 0.01
-
3-phospho-D-glyceroyl phosphate mutant enzyme T33A/S188A, reaction with NADH Spinacia oleracea
1.2.1.13 0.012
-
3-phospho-D-glyceroyl phosphate mutant enzyme S188A, reaction with NADH Spinacia oleracea
1.2.1.13 0.012
-
3-phospho-D-glyceroyl phosphate mutant enzyme T33A, reaction with NADH Spinacia oleracea
1.2.1.13 0.012
-
3-phospho-D-glyceroyl phosphate mutant enzyme T33A/S188A, reaction with NADPH Spinacia oleracea
1.2.1.13 0.012
-
3-phospho-D-glyceroyl phosphate native enzyme A3-GAPDH, reaction with NADH Spinacia oleracea
1.2.1.13 0.013
-
3-phospho-D-glyceroyl phosphate mutant enzyme S188A, reaction with NADPH Spinacia oleracea
1.2.1.13 0.015
-
3-phospho-D-glyceroyl phosphate native enzyme A3-GAPDH, reaction with NADPH Spinacia oleracea
1.2.1.13 0.018
-
3-phospho-D-glyceroyl phosphate mutant enzyme T33A, reaction with NADPH Spinacia oleracea
1.2.1.13 0.029
-
NADPH native enzyme A3-GAPDH Spinacia oleracea
1.2.1.13 0.044
-
NADPH mutant enzyme T33A Spinacia oleracea
1.2.1.13 0.05
-
NADPH mutant enzyme S188A Spinacia oleracea
1.2.1.13 0.05
-
NADPH mutant enzyme T33A/S188A Spinacia oleracea
1.2.1.13 0.14
-
NADH native enzyme A3-GAPDH Spinacia oleracea
1.2.1.13 0.168
-
NADH mutant enzyme S188A Spinacia oleracea
1.2.1.13 0.174
-
NADH mutant enzyme T33A Spinacia oleracea
1.2.1.13 0.184
-
NADH mutant enzyme T33A/S188A Spinacia oleracea

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining

Organism

EC Number Organism UniProt Comment Textmining
1.2.1.13 Spinacia oleracea P19866
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.2.1.13 native and recombinant enzyme Spinacia oleracea

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.2.1.13 3-phospho-D-glyceroyl phosphate + NADH
-
Spinacia oleracea D-glyceraldehyde 3-phosphate + phosphate + NAD+
-
?
1.2.1.13 3-phospho-D-glyceroyl phosphate + NADPH
-
Spinacia oleracea D-glyceraldehyde 3-phosphate + phosphate + NADP+
-
?

Synonyms

EC Number Synonyms Comment Organism
1.2.1.13 A4-GAPDH
-
Spinacia oleracea

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.2.1.13 32
-
NADPH mutant enzyme S188A Spinacia oleracea
1.2.1.13 32
-
NADH mutant enzyme S188A Spinacia oleracea
1.2.1.13 37
-
NADH mutant enzyme T33A Spinacia oleracea
1.2.1.13 41
-
NADH native enzyme A3-GAPDH Spinacia oleracea
1.2.1.13 42
-
NADPH mutant enzyme T33A/S188A Spinacia oleracea
1.2.1.13 48
-
NADH mutant enzyme T33A/S188A Spinacia oleracea
1.2.1.13 52
-
NADPH mutant enzyme T33A Spinacia oleracea
1.2.1.13 61
-
NADPH native enzyme A3-GAPDH Spinacia oleracea

Cofactor

EC Number Cofactor Comment Organism Structure
1.2.1.13 NADH the residues Thr33 and Ser188 are involved in NADPH over NADH selectivity Spinacia oleracea
1.2.1.13 NADPH the residues Thr33 and Ser188 are involved in NADPH over NADH selectivity Spinacia oleracea