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Literature summary for 1.2.1.9 extracted from

  • Sparla, F.; Zaffagnini, M.; Wedel, N.; Scheibe, R.; Pupillo, P.; Trost, P.
    Regulation of photosynthetic GAPDH dissected by mutants (2005), Plant Physiol., 138, 2210-2219.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
into the pET29 vector for expression in Escherichia coli BL21DE3 cells Spinacia oleracea

Protein Variants

Protein Variants Comment Organism
A(plusCTE) chimeric mutant for testing the regulatory function of CTE Spinacia oleracea
B(E326Q) site specific mutant of the GAPDH B-subunit Spinacia oleracea
B(minCTE) deletion mutant for testing the regulatory function of CTE Spinacia oleracea
B(R77A) site specific mutant of the GAPDH B-subunit Spinacia oleracea
B(S188A) site specific mutant of the GAPDH B-subunit Spinacia oleracea

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.012
-
3-phospho-D-glyceroyl phosphate mutant B(E362Q) Spinacia oleracea
0.015
-
3-phospho-D-glyceroyl phosphate GapA Spinacia oleracea
0.015
-
3-phospho-D-glyceroyl phosphate GapB, analyzed under reducing conditions Spinacia oleracea
0.016
-
3-phospho-D-glyceroyl phosphate mutant B(R77A) Spinacia oleracea
0.019
-
3-phospho-D-glyceroyl phosphate A(plusCTE) mutant, analyzed under oxidizing conditions Spinacia oleracea
0.019
-
NADPH mutant B(minCTE) Spinacia oleracea
0.02
-
3-phospho-D-glyceroyl phosphate AB-GAPDH, analyzed under reducing conditions Spinacia oleracea
0.02
-
3-phospho-D-glyceroyl phosphate GapB, analyzed under oxidizing conditions Spinacia oleracea
0.021
-
3-phospho-D-glyceroyl phosphate A(plusCTE) mutant, analyzed under reducing conditions Spinacia oleracea
0.022
-
3-phospho-D-glyceroyl phosphate B(minCTE) mutant Spinacia oleracea
0.024
-
3-phospho-D-glyceroyl phosphate AB-GAPDH, analyzed under oxidizing conditions Spinacia oleracea
0.024
-
NADPH mutant B(188)A, analyzed under oxidizing conditions Spinacia oleracea
0.024
-
NADPH mutant B(E362Q) Spinacia oleracea
0.027
-
3-phospho-D-glyceroyl phosphate mutant B(188)A, analyzed under oxidizing conditions Spinacia oleracea
0.029
-
NADPH GapA Spinacia oleracea
0.03
-
NADPH GapB, analyzed under reducing conditions Spinacia oleracea
0.031
-
3-phospho-D-glyceroyl phosphate mutant B(188)A, analyzed under reducing conditions Spinacia oleracea
0.038
-
NADPH GapB, analyzed under oxidizing conditions Spinacia oleracea
0.043
-
NADPH A(plusCTE) mutant, analyzed under oxidizing conditions Spinacia oleracea
0.046
-
NADPH mutant B(188)A, analyzed under reducing conditions Spinacia oleracea
0.05
-
NADPH AB-GAPDH, analyzed under oxidizing conditions Spinacia oleracea
0.05
-
NADPH AB-GAPDH, analyzed under reducing conditions Spinacia oleracea
0.065
-
NADPH A(plusCTE) mutant, analyzed under reducing conditions Spinacia oleracea
0.078
-
NADPH mutant B(R77A) Spinacia oleracea

Localization

Localization Comment Organism GeneOntology No. Textmining
chloroplast
-
Spinacia oleracea 9507
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
132000
-
GapA, estimated by gel filtration Spinacia oleracea
151000
-
B(minCTE) mutant, estimated by gel filtration Spinacia oleracea
189000
-
tetrameric conformation of GAPDH-A2B2 stabilized by NADPH, estimated by gel filtration Spinacia oleracea
232000
-
GapB mutant B(E326Q) in the presence of NADPH, estimated by gel filtration Spinacia oleracea
243000
-
wild-type GapB, tetramer in the presence of NADPH, estimated by gel filtration Spinacia oleracea
247000
-
GapB mutant B(S188A) in the presence of NADPH, estimated by gel filtration Spinacia oleracea
270000
-
A(plusCTE) mutant, tetramer in the presence of NADPH, estimated by gel filtration Spinacia oleracea
278000
-
GapB mutant B(R77A) in the presence of NADPH, estimated by gel filtration Spinacia oleracea
491000
-
wild-type GapB in the presence of NADH, compatible with octameric structure, estimated by gel filtration Spinacia oleracea
520000
-
GapB mutant B(E326Q) in the presence of NADH, compatible with octameric structure, estimated by gel filtration Spinacia oleracea
520000
-
GapB mutant B(R77A) in the presence of NADH, compatible with octameric structure, estimated by gel filtration Spinacia oleracea
553000
-
GapB mutant B(S188A) in the presence of NADH, compatible with octameric structure, estimated by gel filtration Spinacia oleracea
760000
-
oligomer of GAPDH in the presence of NADH, suggesting A8B8 stoichiometry, estimated by gel filtration Spinacia oleracea

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3-phospho-D-glyceroyl phosphate + NADPH + H+ Spinacia oleracea
-
D-glyceraldehyde 3-phosphate + phosphate + NADP+
-
?

Organism

Organism UniProt Comment Textmining
Spinacia oleracea
-
-
-

Purification (Commentary)

Purification (Comment) Organism
using a Q-Sepharose HP and a MonoQ anion-exchange column Spinacia oleracea

Source Tissue

Source Tissue Comment Organism Textmining
leaf
-
Spinacia oleracea
-

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
GAPDH
-
Spinacia oleracea
glyceraldehyde-3-phosphate dehydrogenase
-
Spinacia oleracea

Temperature Optimum [┬░C]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
25
-
activity assay Spinacia oleracea

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.9
-
activity assay Spinacia oleracea

Cofactor

Cofactor Comment Organism Structure
NADPH
-
Spinacia oleracea