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Literature summary extracted from

  • Blanco, J.; Moore, R.A.; Faehnle, C.R.; Viola, R.E.
    Critical catalytic functional groups in the mechanism of aspartate-beta-semialdehyde dehydrogenase (2004), Acta Crystallogr. Sect. D, 60, 1808-1815.
    View publication on PubMed

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.2.1.11 15 mg/ml purified recombinant wild-type enzyme in 10 mM HEPES, pH 7.0, 1 mM EDTA, and 1 mM DTT, enzyme is free or complexed with substrates phosphate and/or asparate-beta-semialdehyde, hanging drop vapour diffusion method, 20°C, against an equal volume of precipitant solution containing 24-28% PEG 3350, 0.2 M ammonium acetate, and 0.1 M Tris-HCl, pH 8.5, soaking of crystals before harvest in 100 mM phosphate and 2 mM aspartate-beta-semialdehyde, crystallization of mutant H277N in 10 mM HEPES, pH 7.0, 1 mM EDTA, and 1 mM DTT, by addition of precipitant solution containing 5 mM NADP+ and 5 mM inhibitor S-methyl-L-cysteine sulfoxide, 22% PEG 3350, 0.2 M ammonium acetate and 0.1 M sodium cacodylate, pH 6.5, X-ray diffraction structure determination and analysis at about 2.0 A resolution Haemophilus influenzae

Protein Variants

EC Number Protein Variants Comment Organism
1.2.1.11 C136S site-directed mutagenesis, active site mutant is nearly inactive due to decrease in nuleophilicity, and also by a change in the orientation of the histidine imidazole ring Haemophilus influenzae
1.2.1.11 H277N site-directed mutagenesis, active site mutant shows 100fold decreased catalytic efficiency compared to the wild-type enzyme, shift in the position of the bound reaction intermediate Haemophilus influenzae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2.1.11 0.2
-
 NADP+ recombinant wild-type enzyme, pH 9.0, 30°C Haemophilus influenzae
1.2.1.11 0.2
-
 L-aspartate-4-semialdehyde recombinant wild-type enzyme, pH 9.0, 30°C Haemophilus influenzae
1.2.1.11 0.5
-
 L-aspartate-4-semialdehyde recombinant mutant H277N, pH 9.0, 30°C Haemophilus influenzae
1.2.1.11 1.1
-
 NADP+ recombinant mutant H277N, pH 9.0, 30°C Haemophilus influenzae
1.2.1.11 1.6
-
 phosphate recombinant wild-type enzyme, pH 9.0, 30°C Haemophilus influenzae
1.2.1.11 2.7
-
 phosphate recombinant mutant H277N, pH 9.0, 30°C Haemophilus influenzae
1.2.1.11 140
-
 cacodylate recombinant wild-type enzyme, pH 9.0, 30°C Haemophilus influenzae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.2.1.11 L-aspartate-4-semialdehyde + phosphate + NADP+ Haemophilus influenzae reductive dephosphorylation in the aspartate biosynthetic pathway L-4-aspartyl phosphate + NADPH
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.2.1.11 Haemophilus influenzae P44801 gene asd
-

Reaction

EC Number Reaction Comment Organism Reaction ID
1.2.1.11 L-aspartate 4-semialdehyde + phosphate + NADP+ = L-4-aspartyl phosphate + NADPH + H+ catalytic mechanism, function of the catalytic nucleophile Cys136 and the acid-base catalytic His277, the latter is also stabilizing the hemithioacetal intermediate Haemophilus influenzae
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.2.1.11 L-aspartate-4-semialdehyde + cacodylate + NADP+ 10% of the activity with phosphate Haemophilus influenzae L-4-aspartyl cacodylate + NADPH
-
 r
1.2.1.11 L-aspartate-4-semialdehyde + phosphate + NADP+ reductive dephosphorylation in the aspartate biosynthetic pathway Haemophilus influenzae L-4-aspartyl phosphate + NADPH
-
 r
1.2.1.11 L-aspartate-4-semialdehyde + phosphate + NADP+ formation of an acyl-enzyme intermediate Haemophilus influenzae L-4-aspartyl phosphate + NADPH
-
 r

Synonyms

EC Number Synonyms Comment Organism
1.2.1.11 aspartate-beta-semialdehyde dehydrogenase
-
 Haemophilus influenzae
1.2.1.11 hiASADH
-
 Haemophilus influenzae

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.2.1.11 30
-
 assay at Haemophilus influenzae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.2.1.11 3.2
-
 NADP+ recombinant mutant H277N, pH 9.0, 30°C Haemophilus influenzae
1.2.1.11 3.2
-
 phosphate recombinant mutant H277N, pH 9.0, 30°C Haemophilus influenzae
1.2.1.11 3.2
-
 L-aspartate-4-semialdehyde recombinant mutant H277N, pH 9.0, 30°C Haemophilus influenzae
1.2.1.11 330
-
 NADP+ recombinant wild-type enzyme, pH 9.0, 30°C Haemophilus influenzae
1.2.1.11 330
-
 phosphate recombinant wild-type enzyme, pH 9.0, 30°C Haemophilus influenzae
1.2.1.11 330
-
 L-aspartate-4-semialdehyde recombinant wild-type enzyme, pH 9.0, 30°C Haemophilus influenzae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.2.1.11 9
-
 assay at Haemophilus influenzae

Cofactor

EC Number Cofactor Comment Organism Structure
1.2.1.11 NADP+
-
 Haemophilus influenzae
1.2.1.11 NADPH
-
 Haemophilus influenzae