Literature summary for 2.7.7.23 extracted from

Edwards, T.; Gardberg, A.; Phan, I.; Zhang, Y.; Staker, B.; Myler, P.; Lorimer, D.
Structure of uridine diphosphate N-acetylglucosamine pyrophosphorylase from Entamoeba histolytica (2015), Acta Crystallogr. Sect. F, 71, 560-565.

Application

Application	Comment	Organism
drug development	the enzyme is a potential drug target	Entamoeba histolytica

Cloned(Commentary)

Cloned (Comment)	Organism
gene CL6EHI_021200, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli BL21(DE3)	Entamoeba histolytica
gene CL6EHI_039830, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli BL21(DE3)	Entamoeba histolytica

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged enzyme, sitting-drop vapor-diffusion method, mixing of 400 nl of 96.3 mg/ml protein in 25 mM HEPES, pH 7.0, 500 mM NaCl, 5% v/v glycerol, 2 mM DTT, 0.025% w/v azide, with 400 nl of crystallization solution containing 0.2 M lithium sulfate, 0.1 M Bis-Tris, pH 5.5, 25% w/v PEG 3350, 16°C, 1 week, X-ray diffraction structure determination and analysis at 1.8 A resolution, molecular replacement using residues 68-407 of human UAP isoform 1, PDB ID 1jv1, as a search model	Entamoeba histolytica
to 1.8 A resolution. UAP exhibits the same three-domain global architecture as other UAPs, it appears to lack three alpha-helices at the N-terminus and contains two amino acids in the allosteric pocket that make it appear more like the human enzyme than that from Trypanosoma brucei	Entamoeba histolytica

Crystallization (Comment)

Organism

purified recombinant His-tagged enzyme, sitting-drop vapor-diffusion method, mixing of 400 nl of 96.3 mg/ml protein in 25 mM HEPES, pH 7.0, 500 mM NaCl, 5% v/v glycerol, 2 mM DTT, 0.025% w/v azide, with 400 nl of crystallization solution containing 0.2 M lithium sulfate, 0.1 M Bis-Tris, pH 5.5, 25% w/v PEG 3350, 16°C, 1 week, X-ray diffraction structure determination and analysis at 1.8 A resolution, molecular replacement using residues 68-407 of human UAP isoform 1, PDB ID 1jv1, as a search model

Entamoeba histolytica

to 1.8 A resolution. UAP exhibits the same three-domain global architecture as other UAPs, it appears to lack three alpha-helices at the N-terminus and contains two amino acids in the allosteric pocket that make it appear more like the human enzyme than that from Trypanosoma brucei

Entamoeba histolytica

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
UTP + N-acetyl-alpha-D-glucosamine 1-phosphate	Entamoeba histolytica	-	diphosphate + UDP-N-acetyl-alpha-D-glucosamine	-	?

Organism

Organism	UniProt	Comment	Textmining
Entamoeba histolytica	C4M036	-	-
Entamoeba histolytica	C4MA87	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli BL21(DE3) by nickel affinity chromatography and gel filtration, the His-tag is cleaved off by His-tagged MBP-3C protease and removed by affinity chromatography together with the protease	Entamoeba histolytica

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
UTP + N-acetyl-alpha-D-glucosamine 1-phosphate	-	Entamoeba histolytica	diphosphate + UDP-N-acetyl-alpha-D-glucosamine	-	?

Synonyms

Synonyms	Comment	Organism
CL6EHI_021200	-	Entamoeba histolytica
CL6EHI_039830	-	Entamoeba histolytica
EnhiA.01126.a	-	Entamoeba histolytica
EnhiA.01126.b	-	Entamoeba histolytica
UAP	-	Entamoeba histolytica
uridine diphosphate N-acetylglucosamine pyrophosphorylase	-	Entamoeba histolytica

General Information

General Information	Comment	Organism
evolution	although Entamoeba histolytica UAP exhibits the same three-domain global architecture as other UAPs, it appears to lack three alpha-helices at the N-terminus and contains two amino acids in the allosteric pocket that make it appear more like the enzyme from the human host than that from the other parasite Trypanosoma brucei	Entamoeba histolytica
metabolism	uridine diphosphate N-acetylglucosamine pyrophosphorylase catalyzes the final step in the synthesis of UDP-GlcNAc, which is involved in cell-wall biogenesis in plants and fungi and in protein glycosylation	Entamoeba histolytica
additional information	the EhUAP binding pocket largely appears consistent with other UAPs and is likely to follow the common UAP mechanism	Entamoeba histolytica