P54716 (GLVA_BACSU) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Maltose-6'-phosphate glucosidase
EC=3.2.1.122

Alternative name(s):
6-phospho-alpha-D-glucosidase
6-phosphoryl-O-alpha-D-glucopyranosyl:phosphoglucohydrolase

Gene names

Name:	glvA
Synonyms:	glv-1, glvG
Ordered Locus Names:	BSU08180

Organism

Bacillus subtilis (strain 168) [Reference proteome] [HAMAP]

Taxonomic identifier

224308 [NCBI]

Taxonomic lineage

cellular organisms › Bacteria › Firmicutes › Bacilli › Bacillales › Bacillaceae › Bacillus › Bacillus subtilis group › Bacillus subtilis › Bacillus subtilis subsp. subtilis

Protein attributes

Sequence length	449 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Hydrolyzes maltose-6'-phosphate and trehalose-6'-phosphate. Is involved in the catabolism of alpha-glycosides accumulated via a phosphoenolpyruvate-dependent maltose phosphotransferase system (PEP-PTS). Is also able to significantly catalyze the hydrolysis of both 6-phospho-alpha- and 6-phospho-beta-glucosides containing activated leaving groups such as p-nitrophenol and does so with retention and inversion, respectively, of the substrate anomeric configuration.
Catalytic activity	Maltose 6'-phosphate + H2O = D-glucose + D-glucose 6-phosphate.
Cofactor	Binds 1 divalent metal ion per subunit. Manganese, iron, cobalt or nickel enhance activity. Ref.6 Binds 1 NAD+ per subunit. Is only active in the presence of the nicotinamide cofactor in its oxidized, and not the reduced NADH, form. Ref.6
Enzyme regulation	Cellobiose-6'-phosphate and 6-phospho-beta-D-glucopyranoside are not substrates but competitive inhibitors of GlvA. Ref.6
Subunit structure	Homotetramer.
Induction	By maltose; repressed by glucose. Ref.4 Ref.6
Miscellaneous	Reaction proceeds via a redox-elimination-addition mechanism consistent with an E1cb-type mechanism. This includes redox steps involving NAD+ and stabilization of intermediates by Mn2+. Because it hydrolyzes 6-phospho-alpha-glucopyranosides without activated leaving groups (such as maltose-6'-phosphate) but not glycosidic linkage of naturally occurring phospho-beta-glucosides such as cellobiose-6'-phosphate nor methyl 6-phospho-beta-D-glucopyranoside, GlvA is certainly more appropriately classified as a 6-phospho-alpha-glucosidase than as a 6-phospho-beta-glucosidase. The ability to hydrolyze beta-glycosidic linkages is exceptional and applies only to activated 6-phospho-beta-D-glucopyranosides.
Sequence similarities	Belongs to the glycosyl hydrolase 4 family.
Biophysicochemical properties	Kinetic parameters: KM=400 µM for maltose-6'-phosphate (at pH 7.5 and 37 degrees Celsius) Ref.6 KM=360 µM for maltose-6'-phosphate (at pH 8.4 and 37 degrees Celsius) KM=610 µM for methyl 6-phospho-alpha-D-glucoside (at pH 7.5 and 37 degrees Celsius) KM=69 µM for phenyl 6-phospho-alpha-D-glucoside (at pH 7.5 and 37 degrees Celsius) KM=52 µM for 4-nitrophenyl 6-phospho-alpha-D-glucoside (at pH 7.5 and 37 degrees Celsius) KM=12 µM for 3,4-dinitrophenyl 6-phospho-alpha-D-glucoside (at pH 7.5 and 37 degrees Celsius) KM=2.9 µM for 3,4-dinitrophenyl 6-phospho-beta-D-glucoside (at pH 7.5 and 37 degrees Celsius) KM=27 µM for 3,5-dichlorophenyl 6-phospho-beta-D-glucoside (at pH 7.5 and 37 degrees Celsius) KM=34 µM for 3-nitrophenyl 6-phospho-beta-D-glucoside (at pH 7.5 and 37 degrees Celsius) pH dependence: Optimum pH is about 7.6-8.4. Stable from pH 4.0 to 10.0.
Mass spectrometry	Molecular mass is 50510 Da from positions 1 - 449. Determined by ESI. Ref.3

Ontologies

Keywords
Biological process	Carbohydrate metabolism
Ligand	Cobalt Iron Manganese Metal-binding NAD Nickel
Molecular function	Glycosidase Hydrolase
Technical term	3D-structure Complete proteome Direct protein sequencing Reference proteome
Gene Ontology (GO)
Biological_process	carbohydrate metabolic process Inferred from electronic annotation. Source: UniProtKB-KW
Molecular_function	maltose-6'-phosphate glucosidase activity Inferred from electronic annotation. Source: EC metal ion binding Inferred from electronic annotation. Source: UniProtKB-KW nucleotide binding Inferred from electronic annotation. Source: InterPro oxidoreductase activity, acting on the CH-OH group of donors, NAD or NADP as acceptor Inferred from electronic annotation. Source: InterPro
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 449

449

Maltose-6'-phosphate glucosidase

PRO_0000169859

Regions

Nucleotide binding

6 – 72

67

NAD

Sites

Active site

172

1

Proton donor

Active site

265

1

Proton acceptor

Metal binding

171

1

Manganese

Metal binding

202

1

Manganese

Binding site

95

1

Substrate

Binding site

149

1

Substrate

Binding site

285

1

Substrate

Site

111

1

Increases basicity of active site Tyr

Experimental info

Mutagenesis

41

1

D → E or G: Loss of activity.

Mutagenesis

111

1

E → D or G: Loss of activity.

Mutagenesis

359

1

E → D or G: Loss of activity.

Secondary structure

1

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

449

Helix Strand Turn

Details...

Beta strand

6 – 11

6

Beta strand

15 – 17

3

Helix

18 – 27

10

Turn

28 – 31

4

Beta strand

34 – 40

7

Helix

44 – 61

18

Beta strand

65 – 71

7

Helix

73 – 77

5

Beta strand

81 – 85

5

Helix

91 – 103

13

Turn

104 – 106

3

Beta strand

111 – 113

3

Helix

114 – 138

25

Beta strand

143 – 146

4

Helix

151 – 161

11

Beta strand

167 – 169

3

Helix

173 – 185

13

Helix

190 – 192

3

Beta strand

193 – 200

8

Beta strand

203 – 211

9

Helix

218 – 228

11

Helix

246 – 255

10

Beta strand

260 – 262

3

Helix

264 – 266

3

Helix

267 – 270

4

Helix

272 – 276

5

Beta strand

281 – 283

3

Helix

285 – 292

8

Turn

293 – 300

8

Helix

301 – 307

7

Turn

319 – 321

3

Helix

322 – 333

12

Beta strand

337 – 344

8

Beta strand

356 – 365

10

Beta strand

368 – 371

4

Helix

379 – 401

23

Helix

404 – 413

10

Helix

420 – 433

14

Turn

434 – 437

4

Sequences

	Sequence		Length	Mass (Da)
	P54716 [UniParc]. Last modified October 1, 1996. Version 1. Checksum: A903F7E41CFEF7AB	FASTA	449	50,514
	10 20 30 40 50 60 MKKKSFSIVI AGGGSTFTPG IVLMLLDHLE EFPIRKLKLY DNDKERQDRI AGACDVFIRE 70 80 90 100 110 120 KAPDIEFAAT TDPEEAFTDV DFVMAHIRVG KYAMRALDEQ IPLKYGVVGQ ETCGPGGIAY 130 140 150 160 170 180 GMRSIGGVLE ILDYMEKYSP DAWMLNYSNP AAIVAEATRR LRPNSKILNI CDMPVGIEDR 190 200 210 220 230 240 MAQILGLSSR KEMKVRYYGL NHFGWWTSIQ DQEGNDLMPK LKEHVSQYGY IPKTEAEAVE 250 260 270 280 290 300 ASWNDTFAKA RDVQAADPDT LPNTYLQYYL FPDDMVKKSN PNHTRANEVM EGREAFIFSQ 310 320 330 340 350 360 CDMITREQSS ENSEIKIDDH ASYIVDLARA IAYNTGERML LIVENNGAIA NFDPTAMVEV 370 380 390 400 410 420 PCIVGSNGPE PITVGTIPQF QKGLMEQQVS VEKLTVEAWA EKSFQKLWQA LILSKTVPNA 430 440 RVARLILEDL VEANKDFWPE LDQSPTRIS

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Determination of a 12 kb nucleotide sequence around the 76 degrees region of the Bacillus subtilis chromosome." Yamamoto H., Uchiyama S., Fajar A.N., Ogasawara N., Sekiguchi J. Microbiology 142:1417-1421(1996) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: 168.
[2]	"The complete genome sequence of the Gram-positive bacterium Bacillus subtilis." Kunst F., Ogasawara N., Moszer I., Albertini A.M., Alloni G., Azevedo V., Bertero M.G., Bessieres P., Bolotin A., Borchert S., Borriss R., Boursier L., Brans A., Braun M., Brignell S.C., Bron S., Brouillet S., Bruschi C.V. , Caldwell B., Capuano V., Carter N.M., Choi S.-K., Codani J.-J., Connerton I.F., Cummings N.J., Daniel R.A., Denizot F., Devine K.M., Duesterhoeft A., Ehrlich S.D., Emmerson P.T., Entian K.-D., Errington J., Fabret C., Ferrari E., Foulger D., Fritz C., Fujita M., Fujita Y., Fuma S., Galizzi A., Galleron N., Ghim S.-Y., Glaser P., Goffeau A., Golightly E.J., Grandi G., Guiseppi G., Guy B.J., Haga K., Haiech J., Harwood C.R., Henaut A., Hilbert H., Holsappel S., Hosono S., Hullo M.-F., Itaya M., Jones L.-M., Joris B., Karamata D., Kasahara Y., Klaerr-Blanchard M., Klein C., Kobayashi Y., Koetter P., Koningstein G., Krogh S., Kumano M., Kurita K., Lapidus A., Lardinois S., Lauber J., Lazarevic V., Lee S.-M., Levine A., Liu H., Masuda S., Mauel C., Medigue C., Medina N., Mellado R.P., Mizuno M., Moestl D., Nakai S., Noback M., Noone D., O'Reilly M., Ogawa K., Ogiwara A., Oudega B., Park S.-H., Parro V., Pohl T.M., Portetelle D., Porwollik S., Prescott A.M., Presecan E., Pujic P., Purnelle B., Rapoport G., Rey M., Reynolds S., Rieger M., Rivolta C., Rocha E., Roche B., Rose M., Sadaie Y., Sato T., Scanlan E., Schleich S., Schroeter R., Scoffone F., Sekiguchi J., Sekowska A., Seror S.J., Serror P., Shin B.-S., Soldo B., Sorokin A., Tacconi E., Takagi T., Takahashi H., Takemaru K., Takeuchi M., Tamakoshi A., Tanaka T., Terpstra P., Tognoni A., Tosato V., Uchiyama S., Vandenbol M., Vannier F., Vassarotti A., Viari A., Wambutt R., Wedler E., Wedler H., Weitzenegger T., Winters P., Wipat A., Yamamoto H., Yamane K., Yasumoto K., Yata K., Yoshida K., Yoshikawa H.-F., Zumstein E., Yoshikawa H., Danchin A. Nature 390:249-256(1997) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: 168.
[3]	"The gene glvA of Bacillus subtilis 168 encodes a metal-requiring, NAD(H)-dependent 6-phospho-alpha-glucosidase. Assignment to family 4 of the glycosylhydrolase superfamily." Thompson J., Pikis A., Ruvinov S.B., Henrissat B., Yamamoto H., Sekiguchi J. J. Biol. Chem. 273:27347-27356(1998) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 1-37, CHARACTERIZATION, MASS SPECTROMETRY, MUTAGENESIS.
[4]	"Regulation of the glv operon in Bacillus subtilis: YfiA (GlvR) is a positive regulator of the operon that is repressed through CcpA and cre." Yamamoto H., Serizawa M., Thompson J., Sekiguchi J. J. Bacteriol. 183:5110-5121(2001) [PubMed] [Europe PMC] [Abstract] Cited for: INDUCTION.
[5]	"Crystallization and preliminary X-ray analysis of the 6-phospho-alpha-glucosidase from Bacillus subtilis." Varrot A., Yamamoto H., Sekiguchi J., Thompson J., Davies G.J. Acta Crystallogr. D 55:1212-1214(1999) [PubMed] [Europe PMC] [Abstract] Cited for: CRYSTALLIZATION.
[6]	"Mechanism of GlvA from Bacillus subtilis: a detailed kinetic analysis of a 6-phospho-alpha-glucosidase from glycoside hydrolase family 4." Yip V.L.Y., Thompson J., Withers S.G. Biochemistry 46:9840-9852(2007) [PubMed] [Europe PMC] [Abstract] Cited for: REACTION MECHANISM, CHARACTERIZATION, COFACTORS, SUBSTRATE SPECIFICITY, ENZYME REGULATION, BIOPHYSICOCHEMICAL PROPERTIES.
[7]	"Novel catalytic mechanism of glycoside hydrolysis based on the structure of an NAD+/Mn2+ -dependent phospho-alpha-glucosidase from Bacillus subtilis." Rajan S.S., Yang X., Collart F., Yip V.L.Y., Withers S.G., Varrot A., Thompson J., Davies G.J., Anderson W.F. Structure 12:1619-1629(2004) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.05 ANGSTROMS) OF COMPLEX WITH NAD(H) AND ALPHA-D-GLUCOSE-6-PHOSPHATE, REACTION MECHANISM.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

D50543 Genomic DNA. Translation: BAA09103.1.
AL009126 Genomic DNA. Translation: CAB12647.1.

PIR

F69635.

RefSeq

NP_388699.1. NC_000964.3.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1U8X	X-ray	2.05	X	1-449	[»]

ProteinModelPortal

P54716.

SMR

P54716. Positions 3-445.

ModBase

Search...

Protein-protein interaction databases

STRING

224308.BSU08180.

Protein family/group databases

CAZy

GH4. Glycoside Hydrolase Family 4.

Proteomic databases

PaxDb

P54716.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

CAB12647; CAB12647; BSU08180.

GeneID

936161.

KEGG

bsu:BSU08180.

PATRIC

18973286. VBIBacSub10457_0857.

Organism-specific databases

GenoList

BSU08180. [Micado]

Phylogenomic databases

eggNOG

COG1486.

HOGENOM

HOG000239810.

KO

K01232.

OMA

DHASYIV.

ProtClustDB

CLSK880767.

Enzyme and pathway databases

BioCyc

BSUB:BSU08180-MONOMER.

BRENDA

3.2.1.122. 700.

SABIO-RK

P54716.

Family and domain databases

Gene3D

3.40.50.720. 1 hit.
3.90.110.10. 1 hit.

InterPro

IPR019802. GlycHydrolase_4_CS.
IPR001088. Glyco_hydro_4.
IPR022616. Glyco_hydro_4_C.
IPR015955. Lactate_DH/Glyco_Ohase_4_C.
IPR016040. NAD(P)-bd_dom.
[Graphical view]

Pfam

PF02056. Glyco_hydro_4. 1 hit.
PF11975. Glyco_hydro_4C. 1 hit.
[Graphical view]

PRINTS

PR00732. GLHYDRLASE4.

SUPFAM

SSF56327. Lactate_DH/Glyco_hydro_4_C. 1 hit.

PROSITE

PS01324. GLYCOSYL_HYDROL_F4. 1 hit.
[Graphical view]

ProtoNet

Search...

Other

EvolutionaryTrace

P54716.

Entry information

Entry name

GLVA_BACSU

Accession

Primary (citable) accession number: P54716

Entry history

Integrated into UniProtKB/Swiss-Prot:	October 1, 1996
Last sequence update:	October 1, 1996
Last modified:	May 1, 2013
This is version 107 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program