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P54716 (GLVA_BACSU) Reviewed, UniProtKB/Swiss-Prot

Last modified February 19, 2014. Version 112. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (4) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended 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, malA
Ordered Locus Names:BSU08180
OrganismBacillus subtilis (strain 168) [Reference proteome] [HAMAP]
Taxonomic identifier224308 [NCBI]
Taxonomic lineageBacteriaFirmicutesBacilliBacillalesBacillaceaeBacillus

Protein attributes

Sequence length449 AA.
Sequence statusComplete.
Protein existenceEvidence 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

Alpha-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

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Chain1 – 449449Maltose-6'-phosphate glucosidase
PRO_0000169859

Regions

Nucleotide binding6 – 7267NAD

Sites

Active site1721Proton donor
Active site2651Proton acceptor
Metal binding1711Manganese
Metal binding2021Manganese
Binding site951Substrate
Binding site1491Substrate
Binding site2851Substrate
Site1111Increases basicity of active site Tyr

Experimental info

Mutagenesis411D → E or G: Loss of activity.
Mutagenesis1111E → D or G: Loss of activity.
Mutagenesis3591E → D or G: Loss of activity.

Secondary structure

..................................................................... 449
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
P54716 [UniParc].

Last modified October 1, 1996. Version 1.
Checksum: A903F7E41CFEF7AB

FASTA44950,514
        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 

« Hide

References

« Hide '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. expand/collapse author list , 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, COFACTOR, 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.
PIRF69635.
RefSeqNP_388699.1. NC_000964.3.

3D structure databases

PDBe
RCSB PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
1U8XX-ray2.05X1-449[»]
ProteinModelPortalP54716.
SMRP54716. Positions 3-445.
ModBaseSearch...
MobiDBSearch...

Protein-protein interaction databases

STRING224308.BSU08180.

Protein family/group databases

CAZyGH4. Glycoside Hydrolase Family 4.

Proteomic databases

PaxDbP54716.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Genome annotation databases

EnsemblBacteriaCAB12647; CAB12647; BSU08180.
GeneID936161.
KEGGbsu:BSU08180.
PATRIC18973286. VBIBacSub10457_0857.

Organism-specific databases

GenoListBSU08180. [Micado]

Phylogenomic databases

eggNOGCOG1486.
HOGENOMHOG000239810.
KOK01232.
OMAIDDHASY.
OrthoDBEOG632D2X.
ProtClustDBCLSK880767.

Enzyme and pathway databases

BioCycBSUB:BSU08180-MONOMER.
BRENDA3.2.1.122. 700.
SABIO-RKP54716.

Family and domain databases

Gene3D3.40.50.720. 1 hit.
3.90.110.10. 1 hit.
InterProIPR019802. 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]
PfamPF02056. Glyco_hydro_4. 1 hit.
PF11975. Glyco_hydro_4C. 1 hit.
[Graphical view]
PRINTSPR00732. GLHYDRLASE4.
SUPFAMSSF56327. SSF56327. 1 hit.
PROSITEPS01324. GLYCOSYL_HYDROL_F4. 1 hit.
[Graphical view]
ProtoNetSearch...

Other

EvolutionaryTraceP54716.

Entry information

Entry nameGLVA_BACSU
AccessionPrimary (citable) accession number: P54716
Entry history
Integrated into UniProtKB/Swiss-Prot: October 1, 1996
Last sequence update: October 1, 1996
Last modified: February 19, 2014
This is version 112 of the entry and version 1 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Relevant documents

SIMILARITY comments

Index of protein domains and families

PDB cross-references

Index of Protein Data Bank (PDB) cross-references

Glycosyl hydrolases

Classification of glycosyl hydrolase families and list of entries

Bacillus subtilis

Bacillus subtilis (strain 168): entries, gene names and cross-references to SubtiList