Literature summary for 1.2.1.11 extracted from

Blanco, J.; Moore, R.A.; Faehnle, C.R.; Coe, D.M.; Viola, R.E.
The role of substrate-binding groups in the mechanism of aspartate-beta-semialdehyde dehydrogenase (2004), Acta Crystallogr. Sect. D, 60, 1388-1395.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutant enzymes	Haemophilus influenzae

Crystallization (Commentary)

Crystallization (Comment)	Organism
10 mg/ml purified recombinant wild-type and mutant enzymes free or in complex with the substrates, protein in 10 mM HEPES, pH 7.0, 1 mM EDTA, 1 mM DTT, by hanging drop vapour diffusion, 20°C, mixed with equal volume of precipitant solution containing 22-24% PEG 3350, and 0.2 M ammonium acetate, crystals are soaked for 1 h in a solution containing 2 mM aspartate-beta-semialdehyde or 100 mM phosphate, 26% PEG 3350, 0.2 M ammonium acetate, 0.1 M Tris-HCl, pH 8.5, and 20% glycerol, X-ray diffraction structure determination and analysis at about 2.0 A resolution	Haemophilus influenzae

Crystallization (Comment)

Organism

10 mg/ml purified recombinant wild-type and mutant enzymes free or in complex with the substrates, protein in 10 mM HEPES, pH 7.0, 1 mM EDTA, 1 mM DTT, by hanging drop vapour diffusion, 20°C, mixed with equal volume of precipitant solution containing 22-24% PEG 3350, and 0.2 M ammonium acetate, crystals are soaked for 1 h in a solution containing 2 mM aspartate-beta-semialdehyde or 100 mM phosphate, 26% PEG 3350, 0.2 M ammonium acetate, 0.1 M Tris-HCl, pH 8.5, and 20% glycerol, X-ray diffraction structure determination and analysis at about 2.0 A resolution

Haemophilus influenzae

Protein Variants

Protein Variants	Comment	Organism
E243D	site-directed mutagenesis, unaltered Km for the substrates but 8fold increased Km for cofactor NADP+, active site structural alterations	Haemophilus influenzae
K246R	site-directed mutagenesis, mutation of a putative phosphate binding residue, unaltered substrate Km, active site structural alterations	Haemophilus influenzae
R103K	site-directed mutagenesis, adversely affected interaction between enzyme and phosphate, active site structural alterations	Haemophilus influenzae
R103L	site-directed mutagenesis, altered interaction between enzyme and phosphate, active site structural alterations	Haemophilus influenzae
R270K	site-directed mutagenesis, active site mutant, unaltered substrate Km, active site structural alterations	Haemophilus influenzae

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.03	-	L-aspartate-4-semialdehyde	recombinant mutant R103L	Haemophilus influenzae
0.1	-	L-aspartate-4-semialdehyde	recombinant mutant R103K and mutant K246L	Haemophilus influenzae
0.11	-	NADP+	recombinant mutant R103L	Haemophilus influenzae
0.17	-	NADP+	recombinant mutant R270K	Haemophilus influenzae
0.2	-	NADP+	recombinant wild-type enzyme	Haemophilus influenzae
0.2	-	L-aspartate-4-semialdehyde	recombinant wild-type enzyme and mutant E243D	Haemophilus influenzae
0.4	-	L-aspartate-4-semialdehyde	recombinant mutant R270K	Haemophilus influenzae
0.6	-	NADP+	recombinant mutant K246L	Haemophilus influenzae
0.7	-	NADP+	recombinant mutant R103K	Haemophilus influenzae
1	-	phosphate	recombinant mutant K246L	Haemophilus influenzae
1.5	-	phosphate	recombinant mutant E243D	Haemophilus influenzae
1.6	-	phosphate	recombinant wild-type enzyme	Haemophilus influenzae
1.6	-	NADP+	recombinant mutant E243D	Haemophilus influenzae
1.9	-	phosphate	recombinant mutant R270K	Haemophilus influenzae
36.6	-	phosphate	recombinant mutant R103K	Haemophilus influenzae
240	-	phosphate	recombinant mutant R103L	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, aspartate-beta-semialdehydr is the key intermediate in biosynthesis of diaminopimelic acid	L-4-aspartyl phosphate + NADPH	-	r

Organism

Organism	UniProt	Comment	Textmining
Haemophilus influenzae	-	gene asd	-

Reaction

Reaction	Comment	Organism	Reaction ID
L-aspartate 4-semialdehyde + phosphate + NADP+ = L-4-aspartyl phosphate + NADPH + H+	catalytic mechanism, role of substrate binding groups, residues Arg270, Glu243, Arg103, and Lys246 are involved	Haemophilus influenzae	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
L-aspartate-4-semialdehyde + phosphate + NADP+	reductive dephosphorylation in the aspartate biosynthetic pathway, aspartate-beta-semialdehydr is the key intermediate in biosynthesis of diaminopimelic acid	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
ASADH	-	Haemophilus influenzae
aspartate-beta-semialdehyde dehydrogenase	-	Haemophilus influenzae

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
additional information	-	stability of recombinant wild-type and mutant enzymes, oveview	Haemophilus influenzae

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.24	-	L-aspartate-4-semialdehyde	recombinant mutant R103L	Haemophilus influenzae
0.4	-	L-aspartate-4-semialdehyde	recombinant mutant R270K	Haemophilus influenzae
1.2	-	L-aspartate-4-semialdehyde	recombinant mutant R103K	Haemophilus influenzae
4	-	L-aspartate-4-semialdehyde	recombinant mutant E243D	Haemophilus influenzae
11	-	L-aspartate-4-semialdehyde	recombinant mutant K246L	Haemophilus influenzae
330	-	L-aspartate-4-semialdehyde	recombinant wild-type enzyme	Haemophilus influenzae

Cofactor

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