P71447 (PGMB_LACLA) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Beta-phosphoglucomutase
Short name=Beta-PGM
EC=5.4.2.6

Gene names

Name:	pgmB
Ordered Locus Names:	LL0429
ORF Names:	L0001

Organism

Lactococcus lactis subsp. lactis (strain IL1403) (Streptococcus lactis) [Reference proteome] [HAMAP]

Taxonomic identifier

272623 [NCBI]

Taxonomic lineage

cellular organisms › Bacteria › Firmicutes › Bacilli › Lactobacillales › Streptococcaceae › Lactococcus › Lactococcus lactis › Lactococcus lactis subsp. lactis

Protein attributes

Sequence length	221 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Catalyzes the interconversion of D-glucose 1-phosphate (G1P) and D-glucose 6-phosphate (G6P), forming beta-D-glucose 1,6-(bis)phosphate (beta-G16P) as an intermediate. The beta-phosphoglucomutase (Beta-PGM) acts on the beta-C1 anomer of G1P. Glucose or lactose are used in preference to maltose, which is only utilized after glucose or lactose has been exhausted. It plays a key role in the regulation of the flow of carbohydrate intermediates in glycolysis and the formation of the sugar nucleotide UDP-glucose. Ref.1 Ref.4
Catalytic activity	Beta-D-glucose 1-phosphate = beta-D-glucose 6-phosphate.
Cofactor	Binds 2 magnesium ions per subunit. Ref.3 Ref.7
Enzyme regulation	Competitively inhibited by alpha-D-galactose-1-phosphate. Ref.8
Subunit structure	Monomer. Ref.3 Ref.5 Ref.8
Subcellular location	Cytoplasm Potential.
Induction	By maltose, trehalose and sucrose and repressed by glucose and lactose. Ref.1 Ref.3 Ref.4 Ref.8
Post-translational modification	Autophosphorylated.
Miscellaneous	The catalysis proceeds via a phosphoenzyme formed by reaction of an active-site nucleophile with the cofactor glucose 1,6-diphosphate (G1,6-diP). The phosphorylated mutase binds either G1P or G6P and transfers the phosphoryl group to the C6OH or C1OH, respectively.
Sequence similarities	Belongs to the HAD-like hydrolase superfamily. CbbY/CbbZ/Gph/YieH family.
Biophysicochemical properties	Kinetic parameters: KM=14.6 µM for beta-glucose 1-phosphate (at pH 7 and 25 degrees Celsius) Ref.4 Ref.7 KM=20 µM for alpha-D-glucose 1,6-bisphosphate (at pH 7 and 25 degrees Celsius) KM=100 µM for alpha-D-fructose 1,6-bisphosphate (at pH 7 and 25 degrees Celsius) KM=270 µM for magnesium (at pH 7 and 25 degrees Celsius) KM=800 µM for acetyl-phosphate (at pH 7 and 25 degrees Celsius) pH dependence: Optimum pH is around 7. Relatively stable in solution within the pH range of 5-9.5.

Ontologies

Keywords
Biological process	Carbohydrate metabolism
Cellular component	Cytoplasm
Ligand	Magnesium Metal-binding
Molecular function	Isomerase
PTM	Phosphoprotein
Technical term	3D-structure Complete proteome Direct protein sequencing Reference proteome
Gene Ontology (GO)
Biological_process	carbohydrate metabolic process Inferred from direct assay Ref.1. Source: UniProtKB
Cellular_component	cytoplasm Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular_function	beta-phosphoglucomutase activity Inferred from direct assay Ref.4. Source: UniProtKB hydrolase activity Inferred from electronic annotation. Source: InterPro magnesium ion binding Inferred from direct assay Ref.3. Source: UniProtKB
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 221

221

Beta-phosphoglucomutase

PRO_0000108053

Regions

Region

8 – 10

3

Substrate binding

Region

44 – 49

6

Substrate binding

Region

114 – 118

5

Substrate binding

Sites

Active site

8

1

Nucleophile

Active site

10

1

Proton donor

Metal binding

8

1

Magnesium 1

Metal binding

8

1

Magnesium 2

Metal binding

10

1

Magnesium 1

Metal binding

10

1

Magnesium 2; via carbonyl oxygen

Metal binding

169

1

Magnesium 1

Metal binding

170

1

Magnesium 1

Metal binding

170

1

Magnesium 2

Binding site

24

1

Substrate

Binding site

52

1

Substrate

Binding site

76

1

Substrate

Binding site

145

1

Substrate

Site

114

1

Important for catalytic activity and assists the phosphoryl transfer reaction to Asp8 by balancing charge and orienting the reacting groups

Site

145

1

Important for catalytic activity and assists the phosphoryl transfer reaction to Asp8 by balancing charge and orienting the reacting groups

Amino acid modifications

Modified residue

8

1

4-aspartylphosphate

Experimental info

Mutagenesis

8

1

D → A: Inactive. Ref.7

Mutagenesis

8

1

D → E: Inactive. Ref.7

Mutagenesis

10

1

D → A: Inactive. Ref.9

Mutagenesis

10

1

D → E: Inactive. Ref.9

Mutagenesis

10

1

D → N: Inactive. Ref.9

Mutagenesis

10

1

D → S: Inactive. Ref.9

Mutagenesis

16

1

T → P: 500-fold reduction in the rate constant for Asp-8 phosphorylation by beta-G1,6bisP. 6,700-fold reduction in the apparent rate constant for cycling of the phosphorylated enzyme to convert beta-G1P to G6P. 13-fold increase in the estimated rate constant for phosphoryl transfer from the phospho-Asp8 to water. Ref.9

Mutagenesis

20

1

H → A: Impairs Asp-8 phosphorylation by beta-G1,6bisP and phosphoryl transfer from the phospho-Asp8 to the substrate beta-G1P. Ref.9

Mutagenesis

20

1

H → N: 300-fold reduction in the conversion of beta-G1P to G6P in the presence of beta-G1,6bisP. Ref.9

Mutagenesis

20

1

H → Q: 8-fold reduction in the conversion of beta-G1P to G6P in the presence of beta-G1,6bisP. Ref.9

Mutagenesis

45

1

K → A: 20'000-fold decrease in Kcat/KM. Ref.4

Mutagenesis

46

1

G → A: 1'000'000-fold decrease in Kcat/KM. Ref.4

Mutagenesis

46

1

G → P: 100'000-fold decrease in Kcat/KM. Ref.4

Mutagenesis

46

1

G → V: 10'000-fold decrease in Kcat/KM. Ref.4

Mutagenesis

49

1

R → K: 1'000'000-fold decrease in Kcat/KM. Ref.4

Mutagenesis

52

1

S → A: Wild-type activity. Ref.4

Mutagenesis

76

1

K → A: 100-fold reduction in the conversion of beta-G1P to G6P in the presence of beta-G1,6bisP. Ref.9

Mutagenesis

170

1

D → A: Impaired, but active with an increase in the affinity for G1P. Ref.7

Sequence conflict

125

1

K → R in CAA94734. Ref.1

Sequence conflict

206

1

Y → H in CAA94734. Ref.1

Secondary structure

1

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221

Helix Strand Turn

Details...

Beta strand

4 – 7

4

Turn

10 – 12

3

Beta strand

13 – 15

3

Helix

16 – 30

15

Helix

38 – 41

4

Turn

42 – 46

5

Helix

49 – 59

11

Beta strand

60 – 62

3

Helix

66 – 83

18

Helix

84 – 86

3

Helix

89 – 91

3

Helix

96 – 105

10

Beta strand

109 – 112

4

Helix

119 – 125

7

Helix

129 – 131

3

Beta strand

133 – 135

3

Turn

138 – 140

3

Beta strand

141 – 143

3

Helix

149 – 157

9

Helix

162 – 164

3

Beta strand

165 – 171

7

Helix

172 – 181

10

Beta strand

184 – 189

6

Helix

191 – 194

4

Beta strand

196 – 203

8

Helix

204 – 206

3

Helix

209 – 217

9

Sequences

	Sequence		Length	Mass (Da)
	P71447 [UniParc]. Last modified April 27, 2001. Version 2. Checksum: 53AC0BF0FA249EFC	FASTA	221	24,209
	10 20 30 40 50 60 MFKAVLFDLD GVITDTAEYH FRAWKALAEE IGINGVDRQF NEQLKGVSRE DSLQKILDLA 70 80 90 100 110 120 DKKVSAEEFK ELAKRKNDNY VKMIQDVSPA DVYPGILQLL KDLRSNKIKI ALASASKNGP 130 140 150 160 170 180 FLLEKMNLTG YFDAIADPAE VAASKPAPDI FIAAAHAVGV APSESIGLED SQAGIQAIKD 190 200 210 220 SGALPIGVGR PEDLGDDIVI VPDTSYYTLE FLKEVWLQKQ K

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Product formation and phosphoglucomutase activities in Lactococcus lactis: cloning and characterization of a novel phosphoglucomutase gene." Qian N., Stanley G.A., Bunte A., Raadstroem P. Microbiology 143:855-865(1997) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-15, FUNCTION AS A PHOSPHOGLUCOMUTASE AND IN THE CARBOHYDRATE METABOLISM, INDUCTION, SUBSTRATE SPECIFICITY, NOMENCLATURE. Strain: ATCC 19435 / DSM 20481 / NCDO 604 / NCIB 6681 / NCTC 6681.
[2]	"The complete genome sequence of the lactic acid bacterium Lactococcus lactis ssp. lactis IL1403." Bolotin A., Wincker P., Mauger S., Jaillon O., Malarme K., Weissenbach J., Ehrlich S.D., Sorokin A. Genome Res. 11:731-753(2001) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: IL1403.
[3]	"Purification and characterization of two phosphoglucomutases from Lactococcus lactis subsp. lactis and their regulation in maltose- and glucose-utilizing cells." Qian N., Stanley G.A., Hahn-Hagerdal B., Radstrom P. J. Bacteriol. 176:5304-5311(1994) [PubMed] [Europe PMC] [Abstract] Cited for: INDUCTION, SUBSTRATE SPECIFICITY, COFACTOR, SUBUNIT.
[4]	"Analysis of the substrate specificity loop of the HAD superfamily cap domain." Lahiri S.D., Zhang G., Dai J., Dunaway-Mariano D., Allen K.N. Biochemistry 43:2812-2820(2004) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION AS A BETA-PHOSPHOGLUCOMUTASE, MUTAGENESIS OF LYS-45; GLY-46; ARG-49 AND SER-52, INDUCTION, BIOPHYSICOCHEMICAL PROPERTIES. Strain: ATCC 19435 / DSM 20481 / NCDO 604 / NCIB 6681 / NCTC 6681.
[5]	"Caught in the act: the structure of phosphorylated beta-phosphoglucomutase from Lactococcus lactis." Lahiri S.D., Zhang G., Dunaway-Mariano D., Allen K.N. Biochemistry 41:8351-8359(2002) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.30 ANGSTROMS) IN COMPLEX WITH MAGNESIUM IONS, REACTION MECAHNISM, SUBUNIT.
[6]	"The pentacovalent phosphorus intermediate of a phosphoryl transfer reaction." Lahiri S.D., Zhang G., Dunaway-Mariano D., Allen K.N. Science 299:2067-2071(2003) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.20 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOGS AND MAGNESIUM IONS.
[7]	"Catalytic cycling in beta-phosphoglucomutase: a kinetic and structural analysis." Zhang G., Dai J., Wang L., Dunaway-Mariano D., Tremblay L.W., Allen K.N. Biochemistry 44:9404-9416(2005) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) IN COMPLEX WITH MAGNESIUM IONS, MUTAGENESIS OF ASP-8 AND ASP-170, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES, REACTION MECHANISM.
[8]	"Chemical confirmation of a pentavalent phosphorane in complex with beta-phosphoglucomutase." Tremblay L.W., Zhang G., Dai J., Dunaway-Mariano D., Allen K.N. J. Am. Chem. Soc. 127:5298-5299(2005) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOGS AND MAGNESIUM IONS, ENZYME REGULATION, SUBUNIT.
[9]	"Analysis of the structural determinants underlying discrimination between substrate and solvent in beta-phosphoglucomutase catalysis." Dai J., Finci L., Zhang C., Lahiri S., Zhang G., Peisach E., Allen K.N., Dunaway-Mariano D. Biochemistry 48:1984-1995(2009) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.70 ANGSTROMS) IN COMPLEX WITH MAGNESIUM IONS, MUTAGENESIS OF ASP-10; THR-16; HIS-20 AND LYS-76, REACTION MECHANISM.
[10]	"The role of strain in enzyme catalysed phosphate transfer." Bowler M.W., Baxter N.J., Webster C.E., Pollard S., Alizadeh T., Hounslow A.M., Cliff M.J., Bermel W., Williams N.H., Hollfelder F., Blackburn G.M., Waltho J.P. Submitted (APR-2009) to the PDB data bank Cited for: X-RAY CRYSTALLOGRAPHY (1.05 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOGS AND MAGNESIUM IONS.
[11]	"Atomic details of near-transition state conformers for enzyme phosphoryl transfer revealed by MgF-3 rather than by phosphoranes." Baxter N.J., Bowler M.W., Alizadeh T., Cliff M.J., Hounslow A.M., Wu B., Berkowitz D.B., Williams N.H., Blackburn G.M., Waltho J.P. Proc. Natl. Acad. Sci. U.S.A. 107:4555-4560(2010) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.30 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOGS AND MAGNESIUM IONS.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

Z70730 Genomic DNA. Translation: CAA94734.1.
AE005176 Genomic DNA. Translation: AAK04527.1.

PIR

E86678.

RefSeq

NP_266585.1. NC_002662.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1LVH	X-ray	2.30	A/B	1-221	[»]
1O03	X-ray	1.40	A	1-221	[»]
1O08	X-ray	1.20	A	1-221	[»]
1Z4N	X-ray	1.97	A/B	1-221	[»]
1Z4O	X-ray	1.90	A/B	1-221	[»]
1ZOL	X-ray	1.90	A	1-221	[»]
2WF5	X-ray	1.30	A	1-221	[»]
2WF6	X-ray	1.40	A	1-221	[»]
2WF7	X-ray	1.05	A	1-221	[»]
2WF8	X-ray	1.20	A	1-221	[»]
2WF9	X-ray	1.40	A	1-221	[»]
2WFA	X-ray	1.65	A	1-221	[»]
2WHE	X-ray	1.55	A	1-221	[»]
3FM9	X-ray	2.70	A	1-221	[»]

ProteinModelPortal

P71447.

SMR

P71447. Positions 1-221.

ModBase

Search...

Protein-protein interaction databases

STRING

272623.L0001.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

AAK04527; AAK04527; L0001.

GeneID

1114041.

KEGG

lla:L0001.

PATRIC

22293074. VBILacLac136773_0467.

Phylogenomic databases

eggNOG

COG0637.

KO

K01838.

OMA

EEIGING.

ProtClustDB

CLSK876745.

Enzyme and pathway databases

BioCyc

LLAC272623:GHSH-459-MONOMER.
MetaCyc:MONOMER-5821.

BRENDA

5.4.2.6. 2903.

SABIO-RK

P71447.

Family and domain databases

Gene3D

1.10.150.240. 1 hit.
3.40.50.1000. 2 hits.

InterPro

IPR010976. B-phosphoglucomutase_hydrolase.
IPR010972. Beta-phosphoglucomutase.
IPR023214. HAD-like_dom.
IPR006402. HAD-SF_hydro_IA_v3.
IPR005833. Haloacid_DH/epoxide_hydro.
IPR023198. PGP_dom2.
[Graphical view]

Pfam

PF00702. Hydrolase. 1 hit.
[Graphical view]

PRINTS

PR00413. HADHALOGNASE.

SUPFAM

SSF56784. HAD-like_dom. 1 hit.

TIGRFAMs

TIGR01990. bPGM. 1 hit.
TIGR01509. HAD-SF-IA-v3. 1 hit.
TIGR02009. PGMB-YQAB-SF. 1 hit.

ProtoNet

Search...

Other

EvolutionaryTrace

P71447.

Entry information

Entry name

PGMB_LACLA

Accession

Primary (citable) accession number: P71447

Entry history

Integrated into UniProtKB/Swiss-Prot:	July 15, 1998
Last sequence update:	April 27, 2001
Last modified:	May 29, 2013
This is version 103 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program