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Literature summary for 1.2.1.13 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
Search for more literature for 1.2.1.13

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment) Organism
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

Protein Variants Comment Organism
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
T33A affinity for NADPH is significantly decreased, turnover-number for NADPH is lowered Spinacia oleracea
T33A/S188A affinity for NADPH is significantly decreased, turnover-number for NADPH is lowered Spinacia oleracea

KM Value [mM]

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

Organism

Organism UniProt Comment Textmining
Spinacia oleracea P19866
-
-

Purification (Commentary)

Purification (Comment) Organism
native and recombinant enzyme Spinacia oleracea

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
A4-GAPDH
-
 Spinacia oleracea

Turnover Number [1/s]

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

Cofactor

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