Literature summary for 4.1.3.3 extracted from

Huynh, N.; Aye, A.; Li, Y.; Yu, H.; Cao, H.; Tiwari, V.K.; Shin, D.W.; Chen, X.; Fisher, A.J.
Structural basis for substrate specificity and mechanism of N-acetyl-D-neuraminic acid lyase from Pasteurella multocida (2013), Biochemistry, 52, 8570-8579.

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)	Pasteurella multocida

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged wild-type enzyme, wild-type enzyme with Schiff base and in complex with pyruvate, mutant K164A enzyme, and mutant K164A in complex with N-acetylneuraminic acid and N-glycolylneuraminic acid, hanging drop vapour diffusion method, crystals of wild-type PmNAL from 21% PEG 1000, 150 mM NaCl, and 100 mM Na2HPO4/KH2PO4, pH 6.2. PmNAL K164A mutant in ligand-free from from 30% PEG 200, 100 mM NaCl, and acetate, pH 4.5. Crystals of the PmNAL K164A mutant bound to ligands are grown in 38% PEG 300, 0.01 M CaCl2, and 0.1 M sodium cacodylate, pH 6.5. The sialic acid concentration was 5 mM, X-ray diffraction structure determination and analysis at 1.75-2.10 A resolution, molecular replacement modeling	Pasteurella multocida

Crystallization (Comment)

Organism

purified recombinant His-tagged wild-type enzyme, wild-type enzyme with Schiff base and in complex with pyruvate, mutant K164A enzyme, and mutant K164A in complex with N-acetylneuraminic acid and N-glycolylneuraminic acid, hanging drop vapour diffusion method, crystals of wild-type PmNAL from 21% PEG 1000, 150 mM NaCl, and 100 mM Na2HPO4/KH2PO4, pH 6.2. PmNAL K164A mutant in ligand-free from from 30% PEG 200, 100 mM NaCl, and acetate, pH 4.5. Crystals of the PmNAL K164A mutant bound to ligands are grown in 38% PEG 300, 0.01 M CaCl2, and 0.1 M sodium cacodylate, pH 6.5. The sialic acid concentration was 5 mM, X-ray diffraction structure determination and analysis at 1.75-2.10 A resolution, molecular replacement modeling

Pasteurella multocida

Protein Variants

Protein Variants	Comment	Organism
K164A	site-directed mutagenesis, determiniation of three structures of the K164A variant: one in the absence of substrates and two binary complexes with N-acetylneuraminic acid and N-glycolylneuraminic acid. Both sialic acids bind to the active site in the open-chain ketone form of the monosaccharide, every hydroxyl group of the linear sugars makes hydrogen bond interactions with the enzyme	Pasteurella multocida

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
33700	-	x * 33700, recombinant His-tagged enzyme, SDS-PAGE	Pasteurella multocida

Organism

Organism	UniProt	Comment	Textmining
Pasteurella multocida	Q9CKB0	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography	Pasteurella multocida

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
N-acetylneuraminate	reversible aldol cleavage of N-acetylneuraminic acid	Pasteurella multocida	N-acetyl-D-mannosamine + pyruvate	-	r

Subunits

Subunits	Comment	Organism
?	x * 33700, recombinant His-tagged enzyme, SDS-PAGE	Pasteurella multocida

Synonyms

Synonyms	Comment	Organism
NAL	-	Pasteurella multocida