Literature summary for 1.2.1.11 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.

Crystallization (Commentary)

Crystallization (Comment)	Organism
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

Crystallization (Comment)

Organism

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

Protein Variants	Comment	Organism
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
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]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Haemophilus influenzae	P44801	gene asd	-

Reaction

Reaction	Comment	Organism	Reaction ID
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)

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

Synonyms

Synonyms	Comment	Organism
aspartate-beta-semialdehyde dehydrogenase	-	Haemophilus influenzae
hiASADH	-	Haemophilus influenzae

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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

Cofactor

Cofactor	Comment	Organism	Structure
NADP+	-	Haemophilus influenzae
NADPH	-	Haemophilus influenzae