DB code: D00417

RLCP classification 3.133.90010.393 : Transfer
3.1147.6010.88 : Transfer
CATH domain 3.90.550.10 : Spore Coat Polysaccharide Biosynthesis Protein SpsA; Chain A Catalytic domain
2.160.10.10 : UDP N-Acetylglucosamine Acyltransferase; domain 1 Catalytic domain
E.C. 2.7.7.23 2.3.1.157
CSA 1hv9
M-CSA 1hv9
MACiE

CATH domain Related DB codes (homologues)
3.90.550.10 : Spore Coat Polysaccharide Biosynthesis Protein SpsA; Chain A S00709 S00465 S00466 D00859 D00860 T00415
2.160.10.10 : UDP N-Acetylglucosamine Acyltransferase; domain 1 D00464 S00167 D00094

Uniprot Enzyme Name
UniprotKB Protein name Synonyms Includes RefSeq Pfam
P0ACC7 Bifunctional protein GlmU
None UDP-N-acetylglucosamine pyrophosphorylase
EC 2.7.7.23
N-acetylglucosamine-1-phosphate uridyltransferase
Glucosamine-1-phosphate N-acetyltransferase
EC 2.3.1.157
NP_418186.1 (Protein)
NC_000913.2 (DNA/RNA sequence)
YP_491699.1 (Protein)
NC_007779.1 (DNA/RNA sequence)
PF00132 (Hexapep)
PF12804 (NTP_transf_3)
[Graphical View]
Q97R46 Bifunctional protein GlmU
None UDP-N-acetylglucosamine pyrophosphorylase
EC 2.7.7.23
N-acetylglucosamine-1-phosphate uridyltransferase
Glucosamine-1-phosphate N-acetyltransferase
EC 2.3.1.157
NP_345467.1 (Protein)
NC_003028.3 (DNA/RNA sequence)
PF00132 (Hexapep)
PF00483 (NTP_transferase)
[Graphical View]

KEGG enzyme name
UDP-N-acetylglucosamine diphosphorylase
(EC 2.7.7.23 )
UDP-N-acetylglucosamine pyrophosphorylase
(EC 2.7.7.23 )
uridine diphosphoacetylglucosamine pyrophosphorylase
(EC 2.7.7.23 )
UTP:2-acetamido-2-deoxy-alpha-D-glucose-1-phosphateuridylyltransferase
(EC 2.7.7.23 )
UDP-GlcNAc pyrophosphorylase
(EC 2.7.7.23 )
GlmU uridylyltransferase
(EC 2.7.7.23 )
Acetylglucosamine 1-phosphate uridylyltransferase
(EC 2.7.7.23 )
UDP-acetylglucosamine pyrophosphorylase
(EC 2.7.7.23 )
uridine diphosphate-N-acetylglucosamine pyrophosphorylase
(EC 2.7.7.23 )
uridine diphosphoacetylglucosamine phosphorylase
(EC 2.7.7.23 )
acetylglucosamine 1-phosphate uridylyltransferase
(EC 2.7.7.23 )
glucosamine-1-phosphate N-acetyltransferase
(EC 2.3.1.157 )

UniprotKB: Accession Number Entry name Activity Subunit Subcellular location Cofactor
Q97R46 GLMU_STRPN Acetyl-CoA + alpha-D-glucosamine 1-phosphate = CoA + N-acetyl-alpha-D-glucosamine 1-phosphate. UTP + N-acetyl-alpha-D-glucosamine 1-phosphate = diphosphate + UDP-N-acetyl-D-glucosamine. Homotrimer. Cytoplasm. Binds 1 magnesium ion per subunit. Can also use calcium ion to a lesser extent.
P0ACC7 GLMU_ECOLI Acetyl-CoA + alpha-D-glucosamine 1-phosphate = CoA + N-acetyl-alpha-D-glucosamine 1-phosphate. UTP + N-acetyl-alpha-D-glucosamine 1-phosphate = diphosphate + UDP-N-acetyl-D-glucosamine. Homotrimer. In vivo forms an hexameric aggregate. Cytoplasm. Binds 1 magnesium ion per subunit. Can also use cobalt ions to a lesser extent.

KEGG Pathways
Map code Pathways E.C.
MAP00530 Aminosugars metabolism 2.7.7.23 2.3.1.157

Compound table
Cofactors Substrates Products Intermediates
KEGG-id C00305 C00075 C04501 C00024 C06156 C00013 C00043 C00010 C04256
E.C. 2.7.7.23
2.7.7.23
2.7.7.23
2.3.1.157
2.3.1.157
2.7.7.23
2.7.7.23
2.3.1.157
2.3.1.157
Compound Magnesium UTP N-Acetyl-alpha-D-glucosamine 1-phosphate acetyl-CoA D-glucosamine 1-phosphate Pyrophosphate UDP-N-acetyl-D-glucosamine CoA N-acetyl-D-glucosamine 1-phosphate
Type divalent metal (Ca2+, Mg2+) amide group,nucleotide amide group,carbohydrate,phosphate group/phosphate ion amine group,carbohydrate,nucleotide ,peptide/protein,sulfide group amine group,carbohydrate,phosphate group/phosphate ion phosphate group/phosphate ion amide group,carbohydrate,nucleotide amine group,carbohydrate,nucleotide ,peptide/protein,sulfhydryl group amide group,carbohydrate,phosphate group/phosphate ion
ChEBI 18420
15713
16446
15351
27625
29888
16264
15346
7125
PubChem 888
6133
440364
444493
6302
188960
1023
21961011
445675
6816
87642
440272
1fwyA01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Bound:UD1 Unbound Unbound
1fwyB01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Bound:UD1 Unbound Unbound
1fxjA01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Analogue:SO4 Unbound Unbound Unbound
1fxjB01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Analogue:SO4 Unbound Unbound Unbound
1hv9A01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Bound:UD1 Unbound Unbound
1hv9B01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Analogue:_CO Unbound Unbound Unbound Unbound Unbound Bound:UD1 Unbound Unbound
1g95A01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1g97A01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Bound:_MG Unbound Unbound Unbound Unbound Unbound Bound:UD1 Unbound Unbound
1hm0A01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1hm0B01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1hm8A01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1hm8B01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1hm9A01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Analogue:_CA Unbound Unbound Unbound Unbound Unbound Bound:UD1 Unbound Unbound
1hm9B01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Analogue:_CA Unbound Unbound Unbound Unbound Unbound Bound:UD1 Unbound Unbound
1fwyA02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1fwyB02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1fxjA02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1fxjB02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1hv9A02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Unbound Bound:COA Unbound
1hv9B02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Unbound Bound:COA Unbound
1g95A02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1g97A02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1hm0A02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1hm0B02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1hm8A02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Bound:ACO Unbound Unbound Unbound Unbound Unbound
1hm8B02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Bound:ACO Unbound Unbound Unbound Unbound Unbound
1hm9A02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Bound:ACO Unbound Unbound Unbound Unbound Unbound
1hm9B02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Bound:ACO Unbound Unbound Unbound Unbound Unbound

Reference for Active-site residues
resource references E.C.
literature [6], [10]

Active-site residues
PDB Catalytic residues Cofactor-binding residues Modified residues Main-chain involved in catalysis Comment
1fwyA01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ARG 18;LYS 25 ASP 105;ASN 227(Magnesium binding) (2.7.7.23)
1fwyB01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ARG 18;LYS 25 ASP 105;ASN 227(Magnesium binding) (2.7.7.23)
1fxjA01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ARG 18;LYS 25 ASP 105;ASN 227(Magnesium binding) (2.7.7.23)
1fxjB01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ARG 18;LYS 25 ASP 105;ASN 227(Magnesium binding) (2.7.7.23)
1hv9A01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ARG 18;LYS 25 ASP 105;ASN 227(Magnesium binding) (2.7.7.23)
1hv9B01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ARG 18;LYS 25 ASP 105;ASN 227(Magnesium binding) (2.7.7.23)
1g95A01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ARG 15;LYS 22 ASP 102;ASN 227(Magnesium binding) (2.7.7.23)
1g97A01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ARG 15;LYS 22 ASP 102;ASN 227(Magnesium binding) (2.7.7.23)
1hm0A01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ARG 15;LYS 22 ASP 102;ASN 227(Magnesium binding) (2.7.7.23)
1hm0B01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ARG 15;LYS 22 ASP 102;ASN 227(Magnesium binding) (2.7.7.23)
1hm8A01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ARG 15;LYS 22 ASP 102;ASN 227(Magnesium binding) (2.7.7.23)
1hm8B01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ARG 15;LYS 22 ASP 102;ASN 227(Magnesium binding) (2.7.7.23)
1hm9A01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ARG 15;LYS 22 ASP 102;ASN 227(Magnesium binding) (2.7.7.23)
1hm9B01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ARG 15;LYS 22 ASP 102;ASN 227(Magnesium binding) (2.7.7.23)
1fwyA02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ; ; (2.3.1.157) active-site truncated
1fwyB02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ; ; (2.3.1.157) active-site truncated
1fxjA02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ; ; (2.3.1.157) active-site truncated
1fxjB02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ; ; (2.3.1.157) active-site truncated
1hv9A02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 349;HIS 363;SER 405 ALA 380 (2.3.1.157)
1hv9B02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 349;HIS 363;SER 405 ALA 380 (2.3.1.157)
1g95A02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 348;HIS 362;SER 404 ALA 379 (2.3.1.157)
1g97A02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 348;HIS 362;SER 404 ALA 379 (2.3.1.157)
1hm0A02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 348;HIS 362;SER 404 ALA 379 (2.3.1.157)
1hm0B02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 348;HIS 362;SER 404 ALA 379 (2.3.1.157)
1hm8A02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 348;HIS 362;SER 404 ALA 379 (2.3.1.157)
1hm8B02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 348;HIS 362;SER 404 ALA 379 (2.3.1.157)
1hm9A02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 348;HIS 362;SER 404 ALA 379 (2.3.1.157)
1hm9B02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain GLU 348;HIS 362;SER 404 ALA 379 (2.3.1.157)

References for Catalytic Mechanism
References Sections No. of steps in catalysis
[6]
p.4100-4102
[7]
Fig.7, p.286 3
[9]
p.11851

References
[1]
Resource
Comments IDENTIFICATION
Medline ID 94012475
PubMed ID 8407787
Journal J Bacteriol
Year 1993
Volume 175
Pages 6150-7
Authors Mengin-Lecreulx D, van Heijenoort J
Title Identification of the glmU gene encoding N-acetylglucosamine-1-phosphate uridyltransferase in Escherichia coli.
Related PDB
Related UniProtKB P0ACC7
[2]
Resource
Comments CHARACTERIZATION
Medline ID 94364959
PubMed ID 8083170
Journal J Bacteriol
Year 1994
Volume 176
Pages 5788-95
Authors Mengin-Lecreulx D, van Heijenoort J
Title 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.
Related PDB
Related UniProtKB P0ACC7
[3]
Resource
Comments
Medline ID
PubMed ID 7653162
Journal Acta Biochim Pol
Year 1995
Volume 42
Pages 55-9
Authors Szumilo H, Szumilo T, Elbein AD
Title Synthesis of 5-IASA-UDP-GlcNAc and its use for the photoaffinity labeling of a novel UDP-GlcNAc pyrophosphorylase.
Related PDB
Related UniProtKB
[4]
Resource
Comments CHARACTERIZATION
Medline ID 96140233
PubMed ID 8555230
Journal Biochemistry
Year 1996
Volume 35
Pages 579-85
Authors Gehring AM, Lees WJ, Mindiola DJ, Walsh CT, Brown ED
Title Acetyltransfer precedes uridylyltransfer in the formation of UDP-N-acetylglucosamine in separable active sites of the bifunctional GlmU protein of Escherichia coli.
Related PDB
Related UniProtKB P0ACC7
[5]
Resource
Comments
Medline ID
PubMed ID 9733680
Journal J Bacteriol
Year 1998
Volume 180
Pages 4799-803
Authors Pompeo F, van Heijenoort J, Mengin-Lecreulx D
Title Probing the role of cysteine residues in glucosamine-1-phosphate acetyltransferase activity of the bifunctional GlmU protein from Escherichia coli: site-directed mutagenesis and characterization of the mutant enzymes.
Related PDB
Related UniProtKB
[6]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 10428949
Journal EMBO J
Year 1999
Volume 18
Pages 4096-107
Authors Brown K, Pompeo F, Dixon S, Mengin-Lecreulx D, Cambillau C, Bourne Y
Title Crystal structure of the bifunctional N-acetylglucosamine 1-phosphate uridyltransferase from Escherichia coli: a paradigm for the related pyrophosphorylase superfamily.
Related PDB 1fwy 1fxj
Related UniProtKB
[7]
Resource
Comments
Medline ID
PubMed ID 11124906
Journal J Mol Biol
Year 2001
Volume 305
Pages 279-89
Authors Kostrewa D, D'Arcy A, Takacs B, Kamber M
Title Crystal structures of Streptococcus pneumoniae N-acetylglucosamine-1-phosphate uridyltransferase, GlmU, in apo form at 2.33 A resolution and in complex with UDP-N-acetylglucosamine and Mg(2+) at 1.96 A resolution.
Related PDB 1g95 1g97
Related UniProtKB
[8]
Resource
Comments
Medline ID
PubMed ID 11084021
Journal J Biol Chem
Year 2001
Volume 276
Pages 3833-9
Authors Pompeo F, Bourne Y, van Heijenoort J, Fassy F, Mengin-Lecreulx D
Title Dissection of the bifunctional Escherichia coli N-acetylglucosamine-1-phosphate uridyltransferase enzyme into autonomously functional domains and evidence that trimerization is absolutely required for glucosamine-1-phosphate acetyltransferase activity and cell growth.
Related PDB
Related UniProtKB
[9]
Resource
Comments
Medline ID
PubMed ID 11118459
Journal J Biol Chem
Year 2001
Volume 276
Pages 11844-51
Authors Sulzenbacher G, Gal L, Peneff C, Fassy F, Bourne Y
Title Crystal structure of Streptococcus pneumoniae N-acetylglucosamine-1-phosphate uridyltransferase bound to acetyl-coenzyme A reveals a novel active site architecture.
Related PDB 1hm0 1hm8 1hm9
Related UniProtKB
[10]
Resource
Comments
Medline ID
PubMed ID 11329257
Journal Biochemistry
Year 2001
Volume 40
Pages 1913-21
Authors Olsen LR, Roderick SL
Title Structure of the Escherichia coli GlmU pyrophosphorylase and acetyltransferase active sites
Related PDB 1hv9
Related UniProtKB
[11]
Resource
Comments
Medline ID
PubMed ID 11173485
Journal Acta Crystallogr D Biol Crystallogr
Year 2001
Volume 57
Pages 296-7
Authors Olsen LR, Tian Y, Roderick SL
Title Purification, crystallization and preliminary X-ray data for Escherichia coli GlmU: a bifunctional acetyltransferase/uridyltransferase.
Related PDB
Related UniProtKB
[12]
Resource
Comments
Medline ID
PubMed ID 12171937
Journal J Biol Chem
Year 2002
Volume 277
Pages 44214-9
Authors Sivaraman J, Sauve V, Matte A, Cygler M
Title Crystal structure of Escherichia coli glucose-1-phosphate thymidylyltransferase (RffH) complexed with dTTP and Mg2+.
Related PDB
Related UniProtKB

Comments
This protein is a bifunctional enzyme, catalyzing two sequential reactions, with two distinct domains. The first domain functions as a uridylyltransferase (EC 2.7.7.23), which catalyzes the second reaction, whilst the second domain serve as a acetyltransferase (EC 2.3.1.157) that catalyzes the first one (see [4]).
(A) Phosphoryl transfer:
Papers [6] & [7] mentioned catalytic mechanism of uridylyltransferase (EC 2.7.7.23). The paper [7] proposed a catalytic mechanism, in which transphosphorylation is assumed to proceed through SN2 mechanism.
In the proposed mechanism, the reaction proceeds as follows:
(A1) A non-esterified phosphate oxygen atom of GlcNAc-1-P makes a nucleophilic attack on the alpha-phosphate group of UTP, forming penta-coordinated phosphorane transition state with the attacking group and leaving group, pyrophosphate.
(A2) Here, the transition state might be stabilized by the positive charge of Mg2+ ion, which is bound to the oxygen atoms from the transferred alpha-phosphate group and attacking phosphate group as well as Asp102 and Asn227 (PDB;1g97), and the sidechain amino group of Lys22 (PDB;1g97). On the other hand, the leaving pyrophosphate can be stabilized by Arg15 (PDB;1g97).
(A3) The transfer reaction results in the inversion of the configuration at the alpha-phosphate group.
(B) Acyl transfer:
Paper [9] proposed a catalytic mechanism for acyltransferase (EC 2.3.1.157). According to the paper [9], the reaction proceeds as follows:
(B1) Glu348, hydrogen bonding to His362, might modulate the activity of His362.
(B2) His362 (PDB;1hm9) function as a general base, which activates the acceptor amine group of glucosamine-1-P, by abstracting a proton from the amine group.
(B3) The amine group can make a nucleophilic attack on the acyl group of acetyl-CoA.
(B4) Moreover, Ser404 as well as mainchain amide of Ala379 stabilizes the negative charge on the thioester carbonyl during the transition state.

Created Updated
2003-07-22 2009-04-03