Literature summary for 2.7.7.23 extracted from

Vithani, N.; Bais, V.; Prakash, B.
GlmU (N-acetylglucosamine-1-phosphate uridyltransferase) bound to three magnesium ions and ATP at the active site (2014), Acta Crystallogr. Sect. F, 70, 703-708.

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified enzyme complexed with ATP and N-acetyl-alpha-D-glucosamine 1-phosphate, soaking apo GlmUMtb crystals in soaking solution consisting of 10% PEG 8000, 100 mM HEPES, pH 7.5, 20 mM MgCl2, 4 mM CoCl2, 10 mM ATP, and 10 mM GlcNAc-1-P at 4°C, the ligand-bound crystals are cryoprotected in a cryosolution consisting of 20% ethylene glycol, 15% PEG 8000, 100 mM HEPES, pH 7.5, 20 mM MgCl2, and 4 mM CoCl2, X-ray diffraction structure determination and analysis at 1.98 A resolution	Mycobacterium tuberculosis

Crystallization (Comment)

Organism

purified enzyme complexed with ATP and N-acetyl-alpha-D-glucosamine 1-phosphate, soaking apo GlmUMtb crystals in soaking solution consisting of 10% PEG 8000, 100 mM HEPES, pH 7.5, 20 mM MgCl2, 4 mM CoCl2, 10 mM ATP, and 10 mM GlcNAc-1-P at 4°C, the ligand-bound crystals are cryoprotected in a cryosolution consisting of 20% ethylene glycol, 15% PEG 8000, 100 mM HEPES, pH 7.5, 20 mM MgCl2, and 4 mM CoCl2, X-ray diffraction structure determination and analysis at 1.98 A resolution

Mycobacterium tuberculosis

Inhibitors

Inhibitors	Comment	Organism	Structure
ATP	the enzyme binds three magnesium ions and ATP at the active site, but shows no activity with ATP. ATP binding results in an inactive pre-catalytic enzymesubstrate complex, where it adopts an unusual conformation such that the reaction cannot be catalyzed	Mycobacterium tuberculosis
additional information	displacement of MgB 2+ from its usual catalytically competent position, as noted in the crystal structure of RNA polymerase in an inactive state, is considered to be a key factor inhibiting the reaction. The entire metal-substrate complex renders the enzyme catalytically inactive	Mycobacterium tuberculosis

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required, the enzyme utilizes two metal ions, MgA 2+ and MgB 2+, to catalyze the uridyltransfer reaction. The enzyme binds three magnesium ions and ATP at the active site. Displacement of MgB2+ from its usual catalytically competent position, as noted in the crystal structure of RNA polymerase in an inactive state, is considered to be a key factor inhibiting the reaction. In GlmUMtb[GlcNAc-1-P:ATP], the third metal ion, MgC2+ is also stabilized by one coordination interaction with Palpha and five coordination interactions with water molecules. Its coordination by Palpha results in the stabilization of an inactive conformation of ATP. Structure-function analysis, overview. The entire metal-substrate complex renders the enzyme catalytically inactive	Mycobacterium tuberculosis

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	Mycobacterium tuberculosis	-	diphosphate + UDP-N-acetyl-alpha-D-glucosamine	-	?

Organism

Organism	UniProt	Comment	Textmining
Mycobacterium tuberculosis	-	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	enzyme GlmU is specific for its natural substrates UTP and N-acetyl-alpha-D-glucosamine 1-phosphate, substrate specificity, overview. No activity with ATP and TTP as well as glucose-1-phosphate and mannose-1-phosphate, very poor activity with GTP, low activity with CTP	Mycobacterium tuberculosis	?	-	?
UTP + N-acetyl-alpha-D-glucosamine 1-phosphate	-	Mycobacterium tuberculosis	diphosphate + UDP-N-acetyl-alpha-D-glucosamine	-	?

Synonyms

Synonyms	Comment	Organism
GlmU	-	Mycobacterium tuberculosis
N-acetylglucosamine-1-phosphate uridyltransferase	-	Mycobacterium tuberculosis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.6	-	assay at	Mycobacterium tuberculosis

General Information

General Information	Comment	Organism
evolution	the N-acetylglucosamine-1-phosphate uridyltransferase (GlmU) is a bifunctional enzyme exclusive to prokaryotes, that belongs to the family of sugar nucleotidyltransferases (SNTs)	Mycobacterium tuberculosis
physiological function	the enzyme binds N-acetylglucosamine-1-phosphate and UTP, and catalyzes an uridyltransfer reaction to synthesize UDP-GlcNAc, an important precursor for cell-wall biosynthesis	Mycobacterium tuberculosis