P0A9C9 (GLPX_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Fructose-1,6-bisphosphatase 1 class 2
Short name=FBPase 1 class 2
EC=3.1.3.11

Alternative name(s):
D-fructose-1,6-bisphosphate 1-phosphohydrolase 1 class 2

Gene names

Name:	glpX
Ordered Locus Names:	b3925, JW3896

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	336 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Catalyzes the hydrolysis of fructose 1,6-bisphosphate to fructose 6-phosphate. Is likely to be involved in gluconeogenesis during growth on glycerol. Also displays a low activity toward glucose 1,6-bisphosphate, and no activity against ribulose 1,5-bisphosphate, fructose 2,6-bisphosphate, or fructose 1-phosphate. Ref.6 Ref.7
Catalytic activity	D-fructose 1,6-bisphosphate + H2O = D-fructose 6-phosphate + phosphate. Ref.6 Ref.7
Cofactor	Manganese. Mg2+, Co2+, Ni2+, Ca2+, Cu2+ and Zn2+ can not support activity. Ref.6 Ref.7
Enzyme regulation	Competitively inhibited by low concentrations of phosphate (IC50 3.0 mM) and is also sensitive to Li+ (IC50 70 mM). Slightly activated by KCl. Ref.7
Pathway	Carbohydrate biosynthesis; gluconeogenesis.
Subunit structure	Homodimer. Ref.6 Ref.7
Subcellular location	Cytoplasm.
Induction	By glycerol and sn-glycerol-3-phosphate. Ref.1 Ref.7
Disruption phenotype	Cells lacking this gene grow normally on gluconeogenic substrates (succinate or glycerol). Ref.6
Miscellaneous	E.coli K12 also possesses a FBPase class 1 (Fbp), which is the primary FBPase in E.coli and probably represents the main gluconeogenic FBPase.
Sequence similarities	Belongs to the FBPase class 2 family.
Biophysicochemical properties	Kinetic parameters: The catalytic efficiency of GlpX is 3-fold higher than that of YggF, the other FBPase class 2 in E.coli. KM=35 µM for fructose 1,6-bisphosphate (at pH 7.7 and room temperature) Ref.6 Ref.7 KM=70 µM for fructose 1,6-bisphosphate (at pH 9.0 and 37 degrees Celsius) KM=0.6 mM for Mn2+ (at pH 9.0 and 37 degrees Celsius) Vmax=3.3 µmol/min/mg enzyme (at pH 7.7 and room temperature) Vmax=8.8 µmol/min/mg enzyme (at pH 9.0 and 37 degrees Celsius) pH dependence: Optimum pH is 7.5-8.
Sequence caution	The sequence M19644 differs from that shown. Reason: The C-terminal part of glpX was incorrectly assigned as being part of mvrA.

Ontologies

Keywords
Biological process	Carbohydrate metabolism
Cellular component	Cytoplasm
Ligand	Manganese Metal-binding
Molecular function	Hydrolase
Technical term	3D-structure Complete proteome Reference proteome
Gene Ontology (GO)
Biological_process	gluconeogenesis Inferred from electronic annotation. Source: UniProtKB-UniPathway glycerol metabolic process Inferred from electronic annotation. Source: InterPro
Cellular_component	cytoplasm Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular_function	fructose 1,6-bisphosphate 1-phosphatase activity Inferred from direct assay Ref.6. Source: EcoCyc manganese 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 – 336

336

Fructose-1,6-bisphosphatase 1 class 2

PRO_0000201100

Regions

Region

88 – 90

3

Substrate binding

Region

164 – 166

3

Substrate binding

Region

186 – 188

3

Substrate binding

Sites

Metal binding

33

1

Manganese 1

Metal binding

57

1

Manganese 1

Metal binding

85

1

Manganese 2

Metal binding

88

1

Manganese 2

Metal binding

213

1

Manganese 2

Binding site

119

1

Substrate

Binding site

210

1

Substrate; via amide nitrogen

Experimental info

Mutagenesis

29

1

K → A: 2.4-fold increase in FBPase activity, and no effect on substrate affinity. Ref.7

Mutagenesis

57

1

E → A: Strong decrease in FBPase activity. Ref.7

Mutagenesis

59

1

E → A: 5.5-fold decrease in FBPase activity, and 1.4-fold decrease in substrate affinity. Ref.7

Mutagenesis

61

1

D → A: Great decrease in FBPase activity. Ref.7

Mutagenesis

85

1

D → A: Great decrease in FBPase activity. Ref.7

Mutagenesis

88

1

E → A: Strong decrease in FBPase activity. Ref.7

Mutagenesis

90

1

T → A: Strong decrease in FBPase activity. Ref.7

Mutagenesis

119

1

Y → A: Strong decrease in FBPase activity. Ref.7

Mutagenesis

164

1

K → A: Strong decrease in FBPase activity. Ref.7

Mutagenesis

166

1

R → A: Strong decrease in FBPase activity. Ref.7

Mutagenesis

186

1

D → A: 5-fold decrease in FBPase activity, and 3-fold decrease in substrate affinity. Ref.7

Mutagenesis

188

1

D → A: Great decrease in FBPase activity. Ref.7

Mutagenesis

213

1

E → A: Great decrease in FBPase activity. Ref.7

Mutagenesis

235

1

R → A: Nearly no effect on FBPase activity, and 3-fold decrease in substrate affinity. Ref.7

Mutagenesis

239

1

K → A: 1.3-fold increase in FBPase activity, and 1.4-fold decrease in substrate affinity. Ref.7

Secondary structure

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336

Helix Strand Turn

Details...

Helix

3 – 5

3

Helix

6 – 21

16

Turn

22 – 25

4

Helix

29 – 44

16

Beta strand

50 – 58

9

Turn

60 – 62

3

Beta strand

64 – 67

4

Beta strand

71 – 73

3

Beta strand

79 – 88

10

Helix

90 – 94

5

Beta strand

101 – 108

8

Beta strand

117 – 125

9

Helix

127 – 129

3

Helix

139 – 150

12

Helix

154 – 156

3

Beta strand

158 – 162

5

Helix

165 – 167

3

Helix

168 – 177

10

Beta strand

180 – 186

7

Helix

189 – 194

6

Beta strand

204 – 210

7

Helix

211 – 224

14

Beta strand

227 – 233

7

Helix

235 – 237

3

Helix

244 – 257

14

Turn

258 – 260

3

Beta strand

265 – 268

4

Helix

269 – 272

4

Beta strand

278 – 286

9

Beta strand

289 – 291

3

Beta strand

295 – 297

3

Beta strand

300 – 309

10

Turn

310 – 313

4

Beta strand

315 – 323

9

Sequences

	Sequence		Length	Mass (Da)
	P0A9C9 [UniParc]. Last modified July 19, 2005. Version 1. Checksum: DEC2A477E5C4062E	FASTA	336	35,852
	10 20 30 40 50 60 MRRELAIEFS RVTESAALAG YKWLGRGDKN TADGAAVNAM RIMLNQVNID GTIVIGEGEI 70 80 90 100 110 120 DEAPMLYIGE KVGTGRGDAV DIAVDPIEGT RMTAMGQANA LAVLAVGDKG CFLNAPDMYM 130 140 150 160 170 180 EKLIVGPGAK GTIDLNLPLA DNLRNVAAAL GKPLSELTVT ILAKPRHDAV IAEMQQLGVR 190 200 210 220 230 240 VFAIPDGDVA ASILTCMPDS EVDVLYGIGG APEGVVSAAV IRALDGDMNG RLLARHDVKG 250 260 270 280 290 300 DNEENRRIGE QELARCKAMG IEAGKVLRLG DMARSDNVIF SATGITKGDL LEGISRKGNI 310 320 330 ATTETLLIRG KSRTIRRIQS IHYLDRKDPE MQVHIL

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Molecular analysis of the glpFKX regions of Escherichia coli and Shigella flexneri." Truniger V., Boos W., Sweet G. J. Bacteriol. 174:6981-6991(1992) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], INDUCTION. Strain: K12 / MC4100 / ATCC 35695 / DSM 6574.
[2]	"Analysis of the Escherichia coli genome. III. DNA sequence of the region from 87.2 to 89.2 minutes." Plunkett G. III, Burland V., Daniels D.L., Blattner F.R. Nucleic Acids Res. 21:3391-3398(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]. 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]	"Isolation and characterization of methyl viologen-sensitive mutants of Escherichia coli K-12." Morimyo M. J. Bacteriol. 170:2136-2142(1988) [PubMed] [Europe PMC] [Abstract] Cited for: PRELIMINARY NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 228-336. Strain: K12.
[6]	"Purification and characterization of glpX-encoded fructose 1, 6-bisphosphatase, a new enzyme of the glycerol 3-phosphate regulon of Escherichia coli." Donahue J.L., Bownas J.L., Niehaus W.G., Larson T.J. J. Bacteriol. 182:5624-5627(2000) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, CATALYTIC ACTIVITY, COFACTOR, SUBUNIT, KINETIC PARAMETERS, DISRUPTION PHENOTYPE. Strain: K12 / MG1655 / ATCC 47076.
[7]	"Structural and biochemical characterization of the type II fructose-1,6-bisphosphatase GlpX from Escherichia coli." Brown G., Singer A., Lunin V.V., Proudfoot M., Skarina T., Flick R., Kochinyan S., Sanishvili R., Joachimiak A., Edwards A.M., Savchenko A., Yakunin A.F. J. Biol. Chem. 284:3784-3792(2009) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.85 ANGSTROMS) OF APOENZYME AND MUTANT ALA-61 IN COMPLEXES WITH SUBSTRATE; METAL IONS AND PHOSPHATE, FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, COFACTOR, ENZYME REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, REACTION MECHANISM, MUTAGENESIS OF LYS-29; GLU-57; GLU-59; ASP-61; ASP-85; GLU-88; THR-90; TYR-119; LYS-164; ARG-166; ASP-186; ASP-188; GLU-213; ARG-235 AND LYS-239. Strain: K12 / DH5-alpha.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

Z11767 Genomic DNA. Translation: CAA77814.1.
L19201 Genomic DNA. Translation: AAB03057.1.
U00096 Genomic DNA. Translation: AAC76907.1.
AP009048 Genomic DNA. Translation: BAE77385.1.
M19644 Genomic DNA. No translation available.

PIR

A45248.

RefSeq

NP_418360.1. NC_000913.2.
YP_491526.1. NC_007779.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1NI9	X-ray	2.00	A	1-336	[»]
2R8T	X-ray	2.30	A	1-336	[»]
3BIG	X-ray	1.85	A	1-336	[»]
3BIH	X-ray	2.10	A	1-336	[»]
3D1R	X-ray	1.85	A	1-336	[»]

ProteinModelPortal

P0A9C9.

SMR

P0A9C9. Positions 1-323.

ModBase

Search...

Protein-protein interaction databases

IntAct

P0A9C9. 5 interactions.

STRING

511145.b3925.

Proteomic databases

PaxDb

P0A9C9.

PRIDE

P0A9C9.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

AAC76907; AAC76907; b3925.
BAE77385; BAE77385; BAE77385.

GeneID

12931762.
948424.

KEGG

ecj:Y75_p3262.
eco:b3925.

PATRIC

32123363. VBIEscCol129921_4042.

Organism-specific databases

EchoBASE

EB1479.

EcoGene

EG11517. glpX.

Phylogenomic databases

eggNOG

COG1494.

HOGENOM

HOG000241252.

KO

K02446.

OMA

GTVRFIE.

ProtClustDB

PRK09479.

Enzyme and pathway databases

BioCyc

EcoCyc:EG11517-MONOMER.
ECOL316407:JW3896-MONOMER.
MetaCyc:EG11517-MONOMER.

SABIO-RK

P0A9C9.

UniPathway

UPA00138.

Gene expression databases

Genevestigator

P0A9C9.

Family and domain databases

InterPro

IPR004464. FBPase_class-2/SBPase.
[Graphical view]

Pfam

PF03320. FBPase_glpX. 1 hit.
[Graphical view]

PIRSF

PIRSF004532. GlpX. 1 hit.

TIGRFAMs

TIGR00330. glpX. 1 hit.

ProtoNet

Search...

Other

EvolutionaryTrace

P0A9C9.

Entry information

Entry name

GLPX_ECOLI

Accession

Primary (citable) accession number: P0A9C9
Secondary accession number(s): P11007 , P28860, P28900, Q2M8M1

Entry history

Integrated into UniProtKB/Swiss-Prot:	July 19, 2005
Last sequence update:	July 19, 2005
Last modified:	May 1, 2013
This is version 77 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program