Literature summary for 1.2.1.11 extracted from

Dahal, G.; Viola, R.E.
Structure of a fungal form of aspartate semialdehyde dehydrogenase from Cryptococcus neoformans (2015), Acta Crystallogr. Sect. F, 71, 1365-1371 .

Application

Application	Comment	Organism
drug development	the enzyme is a validated target for antimicrobial drug design	Cryptococcus neoformans var. neoformans

Cloned(Commentary)

Cloned (Comment)	Organism
gene CNA02450, codon-optimized, recombinant expression of C-terminally His-tagged enzyme ASADH in Escherichia coli strain BL21(DE3)	Cryptococcus neoformans var. neoformans

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant C-terminally His-tagged enzyme, hanging drop vapour diffusion method, crystallization screening, mixing of 9 mg/ml protein solution with reservoir solution containing 10% PEG 8000, 6% ethylene glycol, 0.1 M HEPES pH 7.5, in a 2:1 ration, at 20°C, crystals are cocrystallized with 4 mM NADP+, X-ray diffraction structure determination and analysis at 2.6 A resolution, modeling by molecular replacement	Cryptococcus neoformans var. neoformans

Crystallization (Comment)

Organism

purified recombinant C-terminally His-tagged enzyme, hanging drop vapour diffusion method, crystallization screening, mixing of 9 mg/ml protein solution with reservoir solution containing 10% PEG 8000, 6% ethylene glycol, 0.1 M HEPES pH 7.5, in a 2:1 ration, at 20°C, crystals are cocrystallized with 4 mM NADP+, X-ray diffraction structure determination and analysis at 2.6 A resolution, modeling by molecular replacement

Cryptococcus neoformans var. neoformans

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
160000	-	recombinant His-tagged enzyme, gel filtration	Cryptococcus neoformans var. neoformans

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
L-aspartate 4-semialdehyde + phosphate + NADP+	Cryptococcus neoformans var. neoformans	-	L-4-aspartyl phosphate + NADPH + H+	-	r
L-aspartate 4-semialdehyde + phosphate + NADP+	Cryptococcus neoformans var. neoformans ATCC MYA-565	-	L-4-aspartyl phosphate + NADPH + H+	-	r

Organism

Organism	UniProt	Comment	Textmining
Cryptococcus neoformans var. neoformans	Q5KPK7	serotype D	-
Cryptococcus neoformans var. neoformans ATCC MYA-565	Q5KPK7	serotype D	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant C-terminally His-tagged enzyme ASADH from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and ultrafiltration	Cryptococcus neoformans var. neoformans

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
L-aspartate 4-semialdehyde + phosphate + NADP+	-	Cryptococcus neoformans var. neoformans	L-4-aspartyl phosphate + NADPH + H+	-	r
L-aspartate 4-semialdehyde + phosphate + NADP+	-	Cryptococcus neoformans var. neoformans ATCC MYA-565	L-4-aspartyl phosphate + NADPH + H+	-	r

Subunits

Subunits	Comment	Organism
More	enzyme secondary structure, crystal structure analysis and comparisons, the C-terminal domain consists of six beta-strands, six alpha-helices and a single 310-helix that represent the catalytic site and the dimerization interface, detailed overview	Cryptococcus neoformans var. neoformans
tetramer	dimer of dimers, 4 * 40000, about, recombinant His-tagged enzyme, SDS-PAGE	Cryptococcus neoformans var. neoformans

Synonyms

Synonyms	Comment	Organism
ASADH	-	Cryptococcus neoformans var. neoformans
CNA02450	-	Cryptococcus neoformans var. neoformans

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8.6	-	assay at	Cryptococcus neoformans var. neoformans

Cofactor

Cofactor	Comment	Organism	Structure
NADP+	the NADP+-binding domain consists of mixed beta-strands flanked on both sides with alpha-helices, with seven beta-strands, five alpha-helices and two 310-helices	Cryptococcus neoformans var. neoformans
NADPH	-	Cryptococcus neoformans var. neoformans

General Information

General Information	Comment	Organism
metabolism	aspartate-semialdehyde dehydrogenase catalyzes the reductive dephosphorylation of the substrate beta-aspartyl phosphate into aspartate semialdehyde, a key intermediate in the aspartate biosynthetic pathway and functions at a critical junction in the aspartate biosynthetic pathway	Cryptococcus neoformans var. neoformans
additional information	The structure of CnASADH belongs to the Rossmann-fold superfamily of pyridine-linked dehydrogenases and shares the same overall monomeric structural features as the other ASADHs for which structures are determined	Cryptococcus neoformans var. neoformans
physiological function	the enzyme catalyzes the NADPH-dependent reductive dephosphorylation of 4-aspartyl phosphate to produce the key intermediate aspartate semialdehyde	Cryptococcus neoformans var. neoformans