P0ACC7 (GLMU_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Bifunctional protein GlmU

Including the following 2 domains:

1. UDP-N-acetylglucosamine pyrophosphorylase
   EC=2.7.7.23
   Alternative name(s):
   N-acetylglucosamine-1-phosphate uridyltransferase
2. Glucosamine-1-phosphate N-acetyltransferase
   EC=2.3.1.157

Gene names

Name:	glmU
Synonyms:	yieA
Ordered Locus Names:	b3730, JW3708

Organism

Escherichia coli (strain K12) [Reference proteome] [HAMAP]

Taxonomic identifier

83333 [NCBI]

Taxonomic lineage

cellular organisms › Bacteria › Proteobacteria › Gammaproteobacteria › Enterobacteriales › Enterobacteriaceae › Escherichia › Escherichia coli

Protein attributes

Sequence length	456 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Catalyzes the last two sequential reactions in the de novo biosynthetic pathway for UDP-N-acetylglucosamine (UDP-GlcNAc). The C-terminal domain catalyzes the transfer of acetyl group from acetyl coenzyme A to glucosamine-1-phosphate (GlcN-1-P) to produce N-acetylglucosamine-1-phosphate (GlcNAc-1-P), which is converted into UDP-GlcNAc by the transfer of uridine 5-monophosphate (from uridine 5-triphosphate), a reaction catalyzed by the N-terminal domain. Ref.6 Ref.7
Catalytic activity	Acetyl-CoA + alpha-D-glucosamine 1-phosphate = CoA + N-acetyl-alpha-D-glucosamine 1-phosphate. Ref.7 UTP + N-acetyl-alpha-D-glucosamine 1-phosphate = diphosphate + UDP-N-acetyl-alpha-D-glucosamine. Ref.7
Cofactor	Binds 1 magnesium ion per subunit. Can also use cobalt ions to a lesser extent. Ref.10
Enzyme regulation	Inhibited by its reaction product N-acetylglucosamine-1-phosphate and by UDP-N-acetylmuramic acid, which is one of the first precursors specific for the peptidoglycan pathway. Ref.6 Ref.12 Ref.13
Pathway	Nucleotide-sugar biosynthesis; UDP-N-acetyl-alpha-D-glucosamine biosynthesis; N-acetyl-alpha-D-glucosamine 1-phosphate from alpha-D-glucosamine 6-phosphate (route II): step 2/2. HAMAP-Rule MF_01631 Nucleotide-sugar biosynthesis; UDP-N-acetyl-alpha-D-glucosamine biosynthesis; UDP-N-acetyl-alpha-D-glucosamine from N-acetyl-alpha-D-glucosamine 1-phosphate: step 1/1. Bacterial outer membrane biogenesis; LPS lipid A biosynthesis. HAMAP-Rule MF_01631
Subunit structure	Homotrimer. In vivo forms a hexameric aggregate. Ref.10 Ref.11 Ref.12 Ref.13
Subcellular location	Cytoplasm HAMAP-Rule MF_01631.
Sequence similarities	In the N-terminal section; belongs to the N-acetylglucosamine-1-phosphate uridyltransferase family. In the C-terminal section; belongs to the transferase hexapeptide repeat family.
Biophysicochemical properties	Kinetic parameters: KM=0.018 mM for N-acetylglucosamine 1-phosphate Ref.6 Ref.7 Ref.8 KM=0.15 mM for alpha-D-glucosamine 1-phosphate KM=0.25 mM for alpha-D-glucosamine 1-phosphate KM=0.32 mM for acetyl-CoA KM=0.6 mM for acetyl-CoA KM=0.84 mM for n-propionyl-CoA KM=1.3 mM for UDP-glucosamine KM=2 mM for glucose-1-P KM=3.7 mM for N-acetylgalactosamine-1-P KM=15 mM for galactosamine-1-phosphate
Sequence caution	The sequence AAA62081.1 differs from that shown. Reason: Frameshift at position 179. Produces two ORFs. The sequence AAA62082.1 differs from that shown. Reason: Frameshift at position 179. Produces two ORFs.

Ontologies

Keywords
Biological process	Cell shape Cell wall biogenesis/degradation Peptidoglycan synthesis
Cellular component	Cytoplasm
Domain	Repeat
Ligand	Cobalt Magnesium Metal-binding
Molecular function	Acyltransferase Nucleotidyltransferase Transferase
Technical term	3D-structure Complete proteome Multifunctional enzyme Reference proteome
Gene Ontology (GO)
Biological_process	UDP-N-acetylglucosamine biosynthetic process Inferred from mutant phenotype Ref.5. Source: EcoCyc cell morphogenesis Inferred from electronic annotation. Source: HAMAP lipid A biosynthetic process Inferred from electronic annotation. Source: UniProtKB-UniPathway lipopolysaccharide biosynthetic process Inferred from electronic annotation. Source: InterPro peptidoglycan biosynthetic process Inferred from electronic annotation. Source: HAMAP regulation of cell shape Inferred from electronic annotation. Source: UniProtKB-KW
Cellular_component	cytoplasm Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular_function	UDP-N-acetylglucosamine diphosphorylase activity Inferred from direct assay Ref.7. Source: EcoCyc glucosamine-1-phosphate N-acetyltransferase activity Inferred from direct assay Ref.6. Source: EcoCyc identical protein binding Inferred from direct assay PubMed 21370307. Source: EcoCyc magnesium ion binding Inferred from direct assay Ref.6. Source: EcoCyc
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 456

456

Bifunctional protein GlmU HAMAP-Rule MF_01631

PRO_0000068701

Regions

Region

1 – 229

229

Pyrophosphorylase HAMAP-Rule MF_01631

Region

11 – 14

4

UDP-GlcNAc binding HAMAP-Rule MF_01631

Region

81 – 82

2

UDP-GlcNAc binding HAMAP-Rule MF_01631

Region

103 – 105

3

UDP-GlcNAc binding HAMAP-Rule MF_01631

Region

230 – 250

21

Linker HAMAP-Rule MF_01631

Region

251 – 456

206

N-acetyltransferase HAMAP-Rule MF_01631

Region

386 – 387

2

Acetyl-CoA binding HAMAP-Rule MF_01631

Sites

Active site

363

1

Proton acceptor By similarity

Metal binding

105

1

Magnesium

Metal binding

227

1

Magnesium

Binding site

25

1

UDP-GlcNAc By similarity

Binding site

76

1

UDP-GlcNAc

Binding site

140

1

UDP-GlcNAc; via amide nitrogen

Binding site

154

1

UDP-GlcNAc

Binding site

169

1

UDP-GlcNAc

Binding site

227

1

UDP-GlcNAc By similarity

Binding site

333

1

UDP-GlcNAc

Binding site

351

1

UDP-GlcNAc

Binding site

366

1

UDP-GlcNAc

Binding site

377

1

UDP-GlcNAc

Binding site

380

1

Acetyl-CoA; via amide nitrogen

Binding site

405

1

Acetyl-CoA

Binding site

423

1

Acetyl-CoA; via amide nitrogen

Binding site

440

1

Acetyl-CoA

Experimental info

Mutagenesis

14

1

G → A: 8-fold decrease in uridyltransferase activity. Creates steric conflict and decreases affinity for UTP. Ref.9

Mutagenesis

18

1

R → A: Dramatically impairs the uridyltransferase activity. Ref.9

Mutagenesis

25

1

K → A: 8-fold decrease in uridyltransferase activity. Ref.9

Mutagenesis

296

1

C → A: No effect. Ref.8

Mutagenesis

307

1

C → A: 1350-fold decrease in acetyltransferase activity. Ref.8

Mutagenesis

324

1

C → A: 8-fold decrease in acetyltransferase activity. Ref.8

Mutagenesis

385

1

C → A: No effect. Ref.8

Sequence conflict

186 – 187

2

KL → NV in CAA25784. Ref.1

Secondary structure

1

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

456

Helix Strand Turn

Details...

Beta strand

6 – 12

7

Helix

17 – 19

3

Helix

25 – 27

3

Beta strand

28 – 30

3

Helix

35 – 46

12

Beta strand

51 – 55

5

Helix

59 – 65

7

Turn

66 – 68

3

Beta strand

71 – 75

5

Helix

82 – 89

8

Helix

90 – 92

3

Beta strand

97 – 103

7

Helix

111 – 120

10

Beta strand

125 – 132

8

Beta strand

141 – 145

5

Beta strand

148 – 153

6

Turn

155 – 157

3

Helix

162 – 164

3

Beta strand

167 – 176

10

Helix

177 – 184

8

Beta strand

191 – 193

3

Helix

198 – 200

3

Helix

201 – 206

6

Turn

207 – 209

3

Beta strand

212 – 215

4

Helix

220 – 223

4

Helix

233 – 249

17

Beta strand

253 – 255

3

Helix

257 – 259

3

Beta strand

260 – 268

9

Beta strand

278 – 286

9

Beta strand

297 – 300

4

Beta strand

314 – 317

4

Beta strand

325 – 327

3

Beta strand

328 – 332

5

Beta strand

336 – 338

3

Beta strand

343 – 355

13

Beta strand

360 – 372

13

Beta strand

383 – 385

3

Beta strand

387 – 390

4

Beta strand

395 – 397

3

Beta strand

408 – 415

8

Sequences

	Sequence		Length	Mass (Da)
	P0ACC7 [UniParc]. Last modified November 8, 2005. Version 1. Checksum: 2F3C5C84F673C8A3	FASTA	456	49,190
	10 20 30 40 50 60 MLNNAMSVVI LAAGKGTRMY SDLPKVLHTL AGKAMVQHVI DAANELGAAH VHLVYGHGGD 70 80 90 100 110 120 LLKQALKDDN LNWVLQAEQL GTGHAMQQAA PFFADDEDIL MLYGDVPLIS VETLQRLRDA 130 140 150 160 170 180 KPQGGIGLLT VKLDDPTGYG RITRENGKVT GIVEHKDATD EQRQIQEINT GILIANGADM 190 200 210 220 230 240 KRWLAKLTNN NAQGEYYITD IIALAYQEGR EIVAVHPQRL SEVEGVNNRL QLSRLERVYQ 250 260 270 280 290 300 SEQAEKLLLA GVMLRDPARF DLRGTLTHGR DVEIDTNVII EGNVTLGHRV KIGTGCVIKN 310 320 330 340 350 360 SVIGDDCEIS PYTVVEDANL AAACTIGPFA RLRPGAELLE GAHVGNFVEM KKARLGKGSK 370 380 390 400 410 420 AGHLTYLGDA EIGDNVNIGA GTITCNYDGA NKFKTIIGDD VFVGSDTQLV APVTVGKGAT 430 440 450 IAAGTTVTRN VGENALAISR VPQTQKEGWR RPVKKK

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"DNA sequence around the Escherichia coli unc operon. Completion of the sequence of a 17 kilobase segment containing asnA, oriC, unc, glmS and phoS." Walker J.E., Gay N.J., Saraste M., Eberle A.N. Biochem. J. 224:799-815(1984) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
[2]	"DNA sequence and analysis of 136 kilobases of the Escherichia coli genome: organizational symmetry around the origin of replication." Burland V.D., Plunkett G. III, Daniels D.L., Blattner F.R. Genomics 16:551-561(1993) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / MG1655 / ATCC 47076.
[3]	"The complete genome sequence of Escherichia coli K-12." Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., Shao Y. Science 277:1453-1474(1997) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA], SEQUENCE REVISION. Strain: K12 / MG1655 / ATCC 47076.
[4]	"Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110." Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T. Mol. Syst. Biol. 2:E1-E5(2006) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[5]	"Identification of the glmU gene encoding N-acetylglucosamine-1-phosphate uridyltransferase in Escherichia coli." Mengin-Lecreulx D., van Heijenoort J. J. Bacteriol. 175:6150-6157(1993) [PubMed] [Europe PMC] [Abstract] Cited for: IDENTIFICATION.
[6]	"Copurification of glucosamine-1-phosphate acetyltransferase and N-acetylglucosamine-1-phosphate uridyltransferase activities of Escherichia coli: characterization of the glmU gene product as a bifunctional enzyme catalyzing two subsequent steps in the pathway for UDP-N-acetylglucosamine synthesis." Mengin-Lecreulx D., van Heijenoort J. J. Bacteriol. 176:5788-5795(1994) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, ENZYME REGULATION.
[7]	"Acetyltransfer precedes uridylyltransfer in the formation of UDP-N-acetylglucosamine in separable active sites of the bifunctional GlmU protein of Escherichia coli." Gehring A.M., Lees W.J., Mindiola D.J., Walsh C.T., Brown E.D. Biochemistry 35:579-585(1996) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL PROPERTIES.
[8]	"Probing the role of cysteine residues in glucosamine-1-phosphate acetyltransferase activity of the bifunctional protein glmU from Escherichia coli: site-directed mutagenesis and characterization of the mutant enzymes." Pompeo F., van Heijenoort J., Mengin-Lecreulx D. J. Bacteriol. 180:4799-4803(1998) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF CYS-296; CYS-307; CYS-324 AND CYS-385, BIOPHYSICOCHEMICAL PROPERTIES.
[9]	"Crystal structure of the bifunctional N-acetylglucosamine 1-phosphate uridyltransferase from Escherichia coli: a paradigm for the related pyrophosphorylase superfamily." Brown K., Pompeo F., Dixon S., Mengin-Lecreulx D., Cambillau C., Bourne Y. EMBO J. 18:4096-4107(1999) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.25 ANGSTROMS) OF THE TRUNCATED FORM AND IN COMPLEX WITH UDP-GLCNAC, MUTAGENESIS OF GLY-14; ARG-18 AND LYS-25.
[10]	"Structure of the Escherichia coli glmU pyrophosphorylase and acetyltransferase active sites." Olsen L.R., Roderick S.L. Biochemistry 40:1913-1921(2001) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS) IN COMPLEX WITH UDP-GLCNAC AND COA, COFACTOR, SUBUNIT.
[11]	"Structure of the E. coli bifunctional GlmU acetyltransferase active site with substrates and products." Olsen L.R., Vetting M.W., Roderick S.L. Protein Sci. 16:1230-1235(2007) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.25 ANGSTROMS) IN COMPLEX WITH UDP-GLCNAC, COA AND MAGNESIUM IONS, SUBUNIT.
[12]	"In vitro validation of acetyltransferase activity of GlmU as an antibacterial target in Haemophilus influenzae." Buurman E.T., Andrews B., Gao N., Hu J., Keating T.A., Lahiri S., Otterbein L.R., Patten A.D., Stokes S.S., Shapiro A.B. J. Biol. Chem. 286:40734-40742(2011) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) OF 233-452, ENZYME REGULATION, SUBUNIT.
[13]	"Inhibitors of acetyltransferase domain of N-acetylglucosamine-1-phosphate-uridyltransferase/glucosamine-1-phosphate-acetyltransferase (GlmU). Part 1: Hit to lead evaluation of a novel arylsulfonamide series." Green O.M., McKenzie A.R., Shapiro A.B., Otterbein L., Ni H., Patten A., Stokes S., Albert R., Kawatkar S., Breed J. Bioorg. Med. Chem. Lett. 22:1510-1519(2012) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) OF 227-456, ENZYME REGULATION, SUBUNIT.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

X01631 Genomic DNA. Translation: CAA25784.1.
L10328 Genomic DNA. Translation: AAA62082.1. Frameshift.
L10328 Genomic DNA. Translation: AAA62081.1. Frameshift.
U00096 Genomic DNA. Translation: AAC76753.1.
AP009048 Genomic DNA. Translation: BAE77558.1.

PIR

C65176.

RefSeq

NP_418186.1. NC_000913.2.
YP_491699.1. NC_007779.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1FWY	X-ray	2.30	A/B	1-331	[»]
1FXJ	X-ray	2.25	A/B	1-331	[»]
1HV9	X-ray	2.10	A/B	1-456	[»]
2OI5	X-ray	2.25	A/B	1-456	[»]
2OI6	X-ray	2.20	A/B	1-456	[»]
2OI7	X-ray	2.54	A/B	1-456	[»]
3TWD	X-ray	1.90	A/B	233-452	[»]
4AA7	X-ray	2.00	A/B	227-456	[»]

ProteinModelPortal

P0ACC7.

SMR

P0ACC7. Positions 3-451.

ModBase

Search...

Protein-protein interaction databases

DIP

DIP-31844N.

IntAct

P0ACC7. 19 interactions.

MINT

MINT-1252604.

STRING

511145.b3730.

Proteomic databases

PaxDb

P0ACC7.

PRIDE

P0ACC7.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

AAC76753; AAC76753; b3730.
BAE77558; BAE77558; BAE77558.

GeneID

12933149.
948246.

KEGG

ecj:Y75_p3438.
eco:b3730.

PATRIC

32122957. VBIEscCol129921_3854.

Organism-specific databases

EchoBASE

EB1184.

EcoGene

EG11198. glmU.

Phylogenomic databases

eggNOG

COG1207.

HOGENOM

HOG000283476.

KO

K04042.

OMA

EPQTHLR.

ProtClustDB

PRK09451.

Enzyme and pathway databases

BioCyc

EcoCyc:NAG1P-URIDYLTRANS-MONOMER.
ECOL316407:JW3708-MONOMER.
MetaCyc:NAG1P-URIDYLTRANS-MONOMER.

UniPathway

UPA00113; UER00532.
UPA00113; UER00533.
UPA00973.

Gene expression databases

Genevestigator

P0ACC7.

Family and domain databases

HAMAP

MF_01631. GlmU.

InterPro

IPR005882. Bifunctional_GlmU.
IPR001451. Hexapep_transf.
IPR018357. Hexapep_transf_CS.
IPR025877. MobA-like_NTP_Trfase_dom.
IPR011004. Trimer_LpxA-like.
[Graphical view]

PANTHER

PTHR22572:SF17. PTHR22572:SF17. 1 hit.

Pfam

PF00132. Hexapep. 3 hits.
PF12804. NTP_transf_3. 1 hit.
[Graphical view]

SUPFAM

SSF51161. Trimer_LpxA_like. 1 hit.

TIGRFAMs

TIGR01173. glmU. 1 hit.

PROSITE

PS00101. HEXAPEP_TRANSFERASES. 1 hit.
[Graphical view]

ProtoNet

Search...

Other

EvolutionaryTrace

P0ACC7.

Entry information

Entry name

GLMU_ECOLI

Accession

Primary (citable) accession number: P0ACC7
Secondary accession number(s): P17114, P76746, Q2M848

Entry history

Integrated into UniProtKB/Swiss-Prot:	November 8, 2005
Last sequence update:	November 8, 2005
Last modified:	May 1, 2013
This is version 81 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program