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Protein

Maltose-6'-phosphate glucosidase

Gene

glvA

Organism
Bacillus subtilis (strain 168)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

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 activityi

Alpha-maltose 6'-phosphate + H2O = D-glucose + D-glucose 6-phosphate.

Cofactori

Protein has several cofactor binding sites:
  • Mn2+1 Publication, Fe2+1 Publication, Co2+1 Publication, Ni2+1 PublicationNote: Binds 1 divalent metal cation per subunit. Manganese, iron, cobalt or nickel enhance activity.1 Publication
  • NAD(+)1 PublicationNote: Binds 1 NAD(+) per subunit. Is only active with NAD(+), not NADH.1 Publication

Enzyme regulationi

Cellobiose-6'-phosphate and 6-phospho-beta-D-glucopyranoside are not substrates but competitive inhibitors of GlvA.1 Publication

Kineticsi

  1. KM=400 µM for maltose-6'-phosphate (at pH 7.5 and 37 degrees Celsius)1 Publication
  2. KM=360 µM for maltose-6'-phosphate (at pH 8.4 and 37 degrees Celsius)1 Publication
  3. KM=610 µM for methyl 6-phospho-alpha-D-glucoside (at pH 7.5 and 37 degrees Celsius)1 Publication
  4. KM=69 µM for phenyl 6-phospho-alpha-D-glucoside (at pH 7.5 and 37 degrees Celsius)1 Publication
  5. KM=52 µM for 4-nitrophenyl 6-phospho-alpha-D-glucoside (at pH 7.5 and 37 degrees Celsius)1 Publication
  6. KM=12 µM for 3,4-dinitrophenyl 6-phospho-alpha-D-glucoside (at pH 7.5 and 37 degrees Celsius)1 Publication
  7. KM=2.9 µM for 3,4-dinitrophenyl 6-phospho-beta-D-glucoside (at pH 7.5 and 37 degrees Celsius)1 Publication
  8. KM=27 µM for 3,5-dichlorophenyl 6-phospho-beta-D-glucoside (at pH 7.5 and 37 degrees Celsius)1 Publication
  9. KM=34 µM for 3-nitrophenyl 6-phospho-beta-D-glucoside (at pH 7.5 and 37 degrees Celsius)1 Publication

    pH dependencei

    Optimum pH is about 7.6-8.4. Stable from pH 4.0 to 10.0.1 Publication

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Binding sitei95 – 951Substrate
    Sitei111 – 1111Increases basicity of active site Tyr
    Binding sitei149 – 1491Substrate
    Metal bindingi171 – 1711Manganese
    Active sitei172 – 1721Proton donor
    Metal bindingi202 – 2021Manganese
    Active sitei265 – 2651Proton acceptor
    Binding sitei285 – 2851Substrate

    Regions

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Nucleotide bindingi6 – 7267NADAdd
    BLAST

    GO - Molecular functioni

    GO - Biological processi

    Complete GO annotation...

    Keywords - Molecular functioni

    Glycosidase, Hydrolase

    Keywords - Biological processi

    Carbohydrate metabolism

    Keywords - Ligandi

    Cobalt, Iron, Manganese, Metal-binding, NAD, Nickel

    Enzyme and pathway databases

    BioCyciBSUB:BSU08180-MONOMER.
    BRENDAi3.2.1.122. 658.
    SABIO-RKP54716.

    Protein family/group databases

    CAZyiGH4. Glycoside Hydrolase Family 4.

    Names & Taxonomyi

    Protein namesi
    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 namesi
    Name:glvA
    Synonyms:glv-1, glvG, malA
    Ordered Locus Names:BSU08180
    OrganismiBacillus subtilis (strain 168)
    Taxonomic identifieri224308 [NCBI]
    Taxonomic lineageiBacteriaFirmicutesBacilliBacillalesBacillaceaeBacillus
    ProteomesiUP000001570 Componenti: Chromosome

    Organism-specific databases

    GenoListiBSU08180. [Micado]

    Pathology & Biotechi

    Mutagenesis

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Mutagenesisi41 – 411D → E or G: Loss of activity. 1 Publication
    Mutagenesisi111 – 1111E → D or G: Loss of activity. 1 Publication
    Mutagenesisi359 – 3591E → D or G: Loss of activity. 1 Publication

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Chaini1 – 449449Maltose-6'-phosphate glucosidasePRO_0000169859Add
    BLAST

    Proteomic databases

    PaxDbiP54716.

    Expressioni

    Inductioni

    By maltose; repressed by glucose.1 Publication

    Interactioni

    Subunit structurei

    Homotetramer.

    Protein-protein interaction databases

    STRINGi224308.Bsubs1_010100004548.

    Structurei

    Secondary structure

    1
    449
    Legend: HelixTurnBeta strand
    Show more details
    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Beta strandi6 – 116Combined sources
    Beta strandi15 – 173Combined sources
    Helixi18 – 2710Combined sources
    Turni28 – 314Combined sources
    Beta strandi34 – 407Combined sources
    Helixi44 – 6118Combined sources
    Beta strandi65 – 717Combined sources
    Helixi73 – 775Combined sources
    Beta strandi81 – 855Combined sources
    Helixi91 – 10313Combined sources
    Turni104 – 1063Combined sources
    Beta strandi111 – 1133Combined sources
    Helixi114 – 13825Combined sources
    Beta strandi143 – 1464Combined sources
    Helixi151 – 16111Combined sources
    Beta strandi167 – 1693Combined sources
    Helixi173 – 18513Combined sources
    Helixi190 – 1923Combined sources
    Beta strandi193 – 2008Combined sources
    Beta strandi203 – 2119Combined sources
    Helixi218 – 22811Combined sources
    Helixi246 – 25510Combined sources
    Beta strandi260 – 2623Combined sources
    Helixi264 – 2663Combined sources
    Helixi267 – 2704Combined sources
    Helixi272 – 2765Combined sources
    Beta strandi281 – 2833Combined sources
    Helixi285 – 2928Combined sources
    Turni293 – 3008Combined sources
    Helixi301 – 3077Combined sources
    Turni319 – 3213Combined sources
    Helixi322 – 33312Combined sources
    Beta strandi337 – 3448Combined sources
    Beta strandi356 – 36510Combined sources
    Beta strandi368 – 3714Combined sources
    Helixi379 – 40123Combined sources
    Helixi404 – 41310Combined sources
    Helixi420 – 43314Combined sources
    Turni434 – 4374Combined sources

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    1U8XX-ray2.05X1-449[»]
    ProteinModelPortaliP54716.
    SMRiP54716. Positions 3-445.
    ModBaseiSearch...
    MobiDBiSearch...

    Miscellaneous databases

    EvolutionaryTraceiP54716.

    Family & Domainsi

    Sequence similaritiesi

    Belongs to the glycosyl hydrolase 4 family.Curated

    Phylogenomic databases

    eggNOGiCOG1486.
    HOGENOMiHOG000239810.
    KOiK01232.
    OMAiKYPMREK.
    OrthoDBiEOG632D2X.
    PhylomeDBiP54716.

    Family and domain databases

    Gene3Di3.40.50.720. 1 hit.
    3.90.110.10. 1 hit.
    InterProiIPR019802. 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]
    PfamiPF02056. Glyco_hydro_4. 1 hit.
    PF11975. Glyco_hydro_4C. 1 hit.
    [Graphical view]
    PRINTSiPR00732. GLHYDRLASE4.
    SUPFAMiSSF56327. SSF56327. 1 hit.
    PROSITEiPS01324. GLYCOSYL_HYDROL_F4. 1 hit.
    [Graphical view]

    Sequencei

    Sequence statusi: Complete.

    P54716-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MKKKSFSIVI AGGGSTFTPG IVLMLLDHLE EFPIRKLKLY DNDKERQDRI
    60 70 80 90 100
    AGACDVFIRE KAPDIEFAAT TDPEEAFTDV DFVMAHIRVG KYAMRALDEQ
    110 120 130 140 150
    IPLKYGVVGQ ETCGPGGIAY GMRSIGGVLE ILDYMEKYSP DAWMLNYSNP
    160 170 180 190 200
    AAIVAEATRR LRPNSKILNI CDMPVGIEDR MAQILGLSSR KEMKVRYYGL
    210 220 230 240 250
    NHFGWWTSIQ DQEGNDLMPK LKEHVSQYGY IPKTEAEAVE ASWNDTFAKA
    260 270 280 290 300
    RDVQAADPDT LPNTYLQYYL FPDDMVKKSN PNHTRANEVM EGREAFIFSQ
    310 320 330 340 350
    CDMITREQSS ENSEIKIDDH ASYIVDLARA IAYNTGERML LIVENNGAIA
    360 370 380 390 400
    NFDPTAMVEV PCIVGSNGPE PITVGTIPQF QKGLMEQQVS VEKLTVEAWA
    410 420 430 440
    EKSFQKLWQA LILSKTVPNA RVARLILEDL VEANKDFWPE LDQSPTRIS
    Length:449
    Mass (Da):50,514
    Last modified:October 1, 1996 - v1
    Checksum:iA903F7E41CFEF7AB
    GO

    Mass spectrometryi

    Molecular mass is 50510 Da from positions 1 - 449. Determined by ESI. 1 Publication

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    D50543 Genomic DNA. Translation: BAA09103.1.
    AL009126 Genomic DNA. Translation: CAB12647.1.
    PIRiF69635.
    RefSeqiNP_388699.1. NC_000964.3.
    WP_003244008.1. NZ_JNCM01000032.1.

    Genome annotation databases

    EnsemblBacteriaiCAB12647; CAB12647; BSU08180.
    GeneIDi936161.
    KEGGibsu:BSU08180.
    PATRICi18973286. VBIBacSub10457_0857.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    D50543 Genomic DNA. Translation: BAA09103.1.
    AL009126 Genomic DNA. Translation: CAB12647.1.
    PIRiF69635.
    RefSeqiNP_388699.1. NC_000964.3.
    WP_003244008.1. NZ_JNCM01000032.1.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    1U8XX-ray2.05X1-449[»]
    ProteinModelPortaliP54716.
    SMRiP54716. Positions 3-445.
    ModBaseiSearch...
    MobiDBiSearch...

    Protein-protein interaction databases

    STRINGi224308.Bsubs1_010100004548.

    Protein family/group databases

    CAZyiGH4. Glycoside Hydrolase Family 4.

    Proteomic databases

    PaxDbiP54716.

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Genome annotation databases

    EnsemblBacteriaiCAB12647; CAB12647; BSU08180.
    GeneIDi936161.
    KEGGibsu:BSU08180.
    PATRICi18973286. VBIBacSub10457_0857.

    Organism-specific databases

    GenoListiBSU08180. [Micado]

    Phylogenomic databases

    eggNOGiCOG1486.
    HOGENOMiHOG000239810.
    KOiK01232.
    OMAiKYPMREK.
    OrthoDBiEOG632D2X.
    PhylomeDBiP54716.

    Enzyme and pathway databases

    BioCyciBSUB:BSU08180-MONOMER.
    BRENDAi3.2.1.122. 658.
    SABIO-RKP54716.

    Miscellaneous databases

    EvolutionaryTraceiP54716.

    Family and domain databases

    Gene3Di3.40.50.720. 1 hit.
    3.90.110.10. 1 hit.
    InterProiIPR019802. 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]
    PfamiPF02056. Glyco_hydro_4. 1 hit.
    PF11975. Glyco_hydro_4C. 1 hit.
    [Graphical view]
    PRINTSiPR00732. GLHYDRLASE4.
    SUPFAMiSSF56327. SSF56327. 1 hit.
    PROSITEiPS01324. GLYCOSYL_HYDROL_F4. 1 hit.
    [Graphical view]
    ProtoNetiSearch...

    Publicationsi

    « Hide 'large scale' publications
    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, 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.

    Entry informationi

    Entry nameiGLVA_BACSU
    AccessioniPrimary (citable) accession number: P54716
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: October 1, 1996
    Last sequence update: October 1, 1996
    Last modified: July 22, 2015
    This is version 122 of the entry and version 1 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    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.

    Keywords - Technical termi

    3D-structure, Complete proteome, Direct protein sequencing, Reference proteome

    Documents

    1. Bacillus subtilis
      Bacillus subtilis (strain 168): entries, gene names and cross-references to SubtiList
    2. Glycosyl hydrolases
      Classification of glycosyl hydrolase families and list of entries
    3. PDB cross-references
      Index of Protein Data Bank (PDB) cross-references
    4. SIMILARITY comments
      Index of protein domains and families

    External Data

    Dasty 3

    Similar proteinsi

    Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
    100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into Uniref entry.
    90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
    50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.