Literature summary for 3.2.1.49 extracted from

Sulzenbacher, G.; Liu, Q.; Bennett, E.; Levery, S.; Bourne, Y.; Ponchel, G.; Clausen, H.; Henrissat, B.
A novel alpha-N-acetylgalactosaminidase family with an NAD+-dependent catalytic mechanism suitable for enzymatic removal of blood group A antigens (2010), Biocatal. Biotransform., 28, 22-32.

No PubMed abstract available

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli	Elizabethkingia meningoseptica

Crystallization (Commentary)

Crystallization (Comment)	Organism
native enzyme and in complex with alpha-N-acetylgalactosamine, to 2.3 and 2.4 A resolution, respectively. Each monomer of NagA consists of two closely associated domains, forming a narrow tunnel in which the NAD+ molecule is anchored. The NAD+ is embedded within a narrow tunnel, almost exclusively shielded from solvent. It is bound with its diphosphate group oriented towards the N-terminus of the first alpha-helix of the N-terminal dinucleotide-binding domain and has both the adenine ring and the nicotinamide ring in the anti conformation. No major structural changes are observed in NagA upon product binding	Elizabethkingia meningoseptica

Crystallization (Comment)

Organism

native enzyme and in complex with alpha-N-acetylgalactosamine, to 2.3 and 2.4 A resolution, respectively. Each monomer of NagA consists of two closely associated domains, forming a narrow tunnel in which the NAD+ molecule is anchored. The NAD+ is embedded within a narrow tunnel, almost exclusively shielded from solvent. It is bound with its diphosphate group oriented towards the N-terminus of the first alpha-helix of the N-terminal dinucleotide-binding domain and has both the adenine ring and the nicotinamide ring in the anti conformation. No major structural changes are observed in NagA upon product binding

Elizabethkingia meningoseptica

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
48500	-	2 * 48500, SDS-PAGE	Elizabethkingia meningoseptica
48500	-	1 * 48500, SDS-PAGE	Elizabethkingia meningoseptica
83000	-	gel filtration	Elizabethkingia meningoseptica

Organism

Organism	UniProt	Comment	Textmining
Elizabethkingia meningoseptica	A4Q8F7	member of glycosyl hydrolase family 109	-

Purification (Commentary)

Purification (Comment)	Organism
-	Elizabethkingia meningoseptica

Reaction

Reaction	Comment	Organism	Reaction ID
alpha-D-GalNAc-(1->3)-beta-D-GalNAc-(1->3)-alpha-D-Gal-(1->4)-beta-D-Gal-(1->4)-beta-D-Glc-(1<->1)-ceramide + H2O = D-GalNAc + beta-D-GalNAc-(1->3)-alpha-D-Gal-(1->4)-beta-D-Gal-(1->4)-beta-D-Glc-(1<->1)-ceramide	a hydride at C3, activated by the interaction of the OH-3 hydroxyl with the conserved His228, is abstracted by NAD+ with oxidation of the OH-3 hydroxyl to a ketone. This activates H2 for proton abstraction by Tyr179, accompanied by elimination of the aglycone, thereby generating a 1,2-unsaturated intermediate. Water is then added to the anomeric center, followed by the reduction of the C3 ketone by the on-board NADH, completing the catalytic sequence and restoring NADH to NAD+ and the enzyme to its starting state to bind another substrate	Elizabethkingia meningoseptica	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
2-nitrophenyl-alpha-N-acetylgalactosaminide + H2O	-	Elizabethkingia meningoseptica	2-nitrophenol + alpha-N-acetylgalactosamine	-	?
additional information	enzyme efficiently cleaves blood group A antigen. Unusual catalytic mechanism involving NAD+. NagA might adopt a mechanism similar to that of glycosyl hydrolase family 4 enzymes utilizing an oxidation-elimination-addition-reduction sequence	Elizabethkingia meningoseptica	?	-	?

Subunits

Subunits	Comment	Organism
dimer	2 * 48500, SDS-PAGE	Elizabethkingia meningoseptica
monomer	1 * 48500, SDS-PAGE	Elizabethkingia meningoseptica
More	enzyme may exist both in the monomeric and dimeric form, depending on the salt concentration of the buffer solution	Elizabethkingia meningoseptica

Synonyms

Synonyms	Comment	Organism
NagA	-	Elizabethkingia meningoseptica

Cofactor

Cofactor	Comment	Organism	Structure
NAD+	binds 1 NAD+ per subunit. The NAD+ cannot dissociate	Elizabethkingia meningoseptica