AGASK_RUMGN
ID AGASK_RUMGN Reviewed; 935 AA.
AC G4T4R7; G3XAP8;
DT 15-FEB-2017, integrated into UniProtKB/Swiss-Prot.
DT 14-DEC-2011, sequence version 1.
DT 03-AUG-2022, entry version 38.
DE RecName: Full=Bifunctional alpha-galactosidase/sucrose kinase AgaSK {ECO:0000303|PubMed:21931163, ECO:0000312|PDB:2YFN, ECO:0000312|PDB:2YFO};
DE Includes:
DE RecName: Full=Alpha-galactosidase {ECO:0000303|PubMed:21931163};
DE EC=3.2.1.22 {ECO:0000269|PubMed:21931163};
DE AltName: Full=Melibiase {ECO:0000305};
DE Includes:
DE RecName: Full=Sucrose kinase {ECO:0000303|PubMed:21931163};
DE EC=2.7.-.- {ECO:0000269|PubMed:21931163};
GN Name=agaSK {ECO:0000303|PubMed:21931163, ECO:0000312|EMBL:CCA61959.1};
GN ORFNames=RUGNEv3_61222 {ECO:0000312|EMBL:CCA61959.1};
OS Ruminococcus gnavus.
OC Bacteria; Firmicutes; Clostridia; Eubacteriales; Lachnospiraceae;
OC Mediterraneibacter.
OX NCBI_TaxID=33038 {ECO:0000312|EMBL:CCA61959.1};
RN [1] {ECO:0007744|PDB:2YFN, ECO:0007744|PDB:2YFO}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], X-RAY CRYSTALLOGRAPHY (1.35 ANGSTROMS)
RP OF 1-720 OF SUBSTRATE-FREE ENZYME AND IN COMPLEX WITH GALACTOSE AND
RP MAGNESIUM, FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND SUBUNIT.
RC STRAIN=E1 {ECO:0000303|PubMed:21931163};
RX PubMed=21931163; DOI=10.1074/jbc.m111.286039;
RA Bruel L., Sulzenbacher G., Cervera Tison M., Pujol A., Nicoletti C.,
RA Perrier J., Galinier A., Ropartz D., Fons M., Pompeo F., Giardina T.;
RT "alpha-Galactosidase/sucrose kinase (AgaSK), a novel bifunctional enzyme
RT from the human microbiome coupling galactosidase and kinase activities.";
RL J. Biol. Chem. 286:40814-40823(2011).
CC -!- FUNCTION: Bifunctional enzyme with alpha-galactosidase and sucrose
CC kinase activities. Produces sucrose-6-phosphate directly from
CC raffinose. Binds ATP. Phosphorylates sucrose specifically on the C6
CC position of glucose in the presence of ATP. Hydrolyzes melibiose,
CC raffinose, stachyose and synthetic substrate p-nitrophenyl-alpha-D-
CC galactopyranoside with high activity. Low activity against locust bean
CC gum, guar gum and synthetic substrates xylose alpha-D-4-nitrophenol,
CC glucose alpha-D-4-nitrophenol and o-nitrophenyl-alpha-D-
CC galactopyranoside. {ECO:0000269|PubMed:21931163}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Hydrolysis of terminal, non-reducing alpha-D-galactose
CC residues in alpha-D-galactosides, including galactose
CC oligosaccharides, galactomannans and galactolipids.; EC=3.2.1.22;
CC Evidence={ECO:0000269|PubMed:21931163};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:21931163};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.37 mM for p-nitrophenyl-alpha-D-galactopyranoside (at pH 6.0 and
CC 37 degrees Celsius) {ECO:0000269|PubMed:21931163};
CC Note=kcat/KM is 1850 s(1) mM(1) for p-nitrophenyl-alpha-D-
CC galactopyranoside. {ECO:0000269|PubMed:21931163};
CC pH dependence:
CC Optimum pH is 6.0. Active between pH 4.0-6.5.
CC {ECO:0000269|PubMed:21931163};
CC Temperature dependence:
CC Optimum temperature is 50 degrees Celsius.
CC {ECO:0000269|PubMed:21931163};
CC -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:21931163}.
CC -!- SIMILARITY: In the N-terminal section; belongs to the glycosyl
CC hydrolase 36 family. {ECO:0000305}.
CC -!- SIMILARITY: In the C-terminal section; belongs to the uridine kinase
CC family. {ECO:0000305}.
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DR EMBL; FQ790379; CCA61959.1; -; Genomic_DNA.
DR PDB; 2YFN; X-ray; 1.45 A; A=1-720.
DR PDB; 2YFO; X-ray; 1.35 A; A=1-720.
DR PDBsum; 2YFN; -.
DR PDBsum; 2YFO; -.
DR AlphaFoldDB; G4T4R7; -.
DR SMR; G4T4R7; -.
DR BioCyc; MetaCyc:MON-18513; -.
DR GO; GO:0004557; F:alpha-galactosidase activity; IDA:UniProtKB.
DR GO; GO:0005524; F:ATP binding; IDA:UniProtKB.
DR GO; GO:0019200; F:carbohydrate kinase activity; IDA:UniProtKB.
DR GO; GO:0000287; F:magnesium ion binding; IDA:UniProtKB.
DR GO; GO:0016779; F:nucleotidyltransferase activity; IDA:UniProtKB.
DR GO; GO:0052692; F:raffinose alpha-galactosidase activity; IDA:UniProtKB.
DR GO; GO:0005995; P:melibiose catabolic process; IDA:UniProtKB.
DR GO; GO:0051289; P:protein homotetramerization; IDA:UniProtKB.
DR GO; GO:0034484; P:raffinose catabolic process; IDA:UniProtKB.
DR GO; GO:0033531; P:stachyose metabolic process; IDA:UniProtKB.
DR CDD; cd14791; GH36; 1.
DR Gene3D; 2.60.40.1180; -; 1.
DR Gene3D; 2.70.98.60; -; 1.
DR Gene3D; 3.20.20.70; -; 1.
DR Gene3D; 3.40.50.300; -; 1.
DR InterPro; IPR013785; Aldolase_TIM.
DR InterPro; IPR038417; Alpga-gal_N_sf.
DR InterPro; IPR000111; Glyco_hydro_27/36_CS.
DR InterPro; IPR002252; Glyco_hydro_36.
DR InterPro; IPR031705; Glyco_hydro_36_C.
DR InterPro; IPR031704; Glyco_hydro_36_N.
DR InterPro; IPR013780; Glyco_hydro_b.
DR InterPro; IPR017853; Glycoside_hydrolase_SF.
DR InterPro; IPR027417; P-loop_NTPase.
DR InterPro; IPR006083; PRK/URK.
DR Pfam; PF16874; Glyco_hydro_36C; 1.
DR Pfam; PF16875; Glyco_hydro_36N; 1.
DR Pfam; PF00485; PRK; 1.
DR PRINTS; PR00743; GLHYDRLASE36.
DR SUPFAM; SSF51445; SSF51445; 1.
DR SUPFAM; SSF52540; SSF52540; 1.
DR PROSITE; PS00512; ALPHA_GALACTOSIDASE; 1.
PE 1: Evidence at protein level;
KW 3D-structure; ATP-binding; Glycosidase; Hydrolase; Kinase; Magnesium;
KW Metal-binding; Multifunctional enzyme; Nucleotide-binding;
KW Nucleotidyltransferase; Transferase.
FT CHAIN 1..935
FT /note="Bifunctional alpha-galactosidase/sucrose kinase
FT AgaSK"
FT /id="PRO_0000438979"
FT REGION 1..720
FT /note="Alpha-galactosidase"
FT /evidence="ECO:0000305"
FT REGION 721..935
FT /note="Sucrose kinase"
FT /evidence="ECO:0000305"
FT ACT_SITE 526
FT /note="Nucleophile"
FT /evidence="ECO:0000250|UniProtKB:Q8A6L0"
FT ACT_SITE 606
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000250|UniProtKB:Q8A6L0"
FT BINDING 176
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:21931163,
FT ECO:0007744|PDB:2YFN"
FT BINDING 277
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:21931163,
FT ECO:0007744|PDB:2YFN"
FT BINDING 280
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000269|PubMed:21931163,
FT ECO:0007744|PDB:2YFN"
FT BINDING 366..367
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21931163,
FT ECO:0007744|PDB:2YFO"
FT BINDING 443
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21931163,
FT ECO:0007744|PDB:2YFO"
FT BINDING 476..480
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21931163,
FT ECO:0007744|PDB:2YFO"
FT BINDING 518..521
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21931163,
FT ECO:0007744|PDB:2YFO"
FT BINDING 540
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21931163,
FT ECO:0007744|PDB:2YFO"
FT BINDING 748..752
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000250|UniProtKB:O43252"
FT BINDING 824
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000250|UniProtKB:O43252"
FT STRAND 3..6
FT /evidence="ECO:0007829|PDB:2YFO"
FT TURN 7..10
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 11..15
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 17..25
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 31..38
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 44..47
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 60..62
FT /evidence="ECO:0007829|PDB:2YFO"
FT TURN 70..72
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 75..77
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 89..93
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 104..113
FT /evidence="ECO:0007829|PDB:2YFO"
FT TURN 127..129
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 130..139
FT /evidence="ECO:0007829|PDB:2YFO"
FT TURN 140..143
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 144..153
FT /evidence="ECO:0007829|PDB:2YFO"
FT TURN 154..157
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 158..167
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 169..171
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 173..187
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 190..198
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 201..209
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 212..219
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 221..224
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 226..228
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 230..236
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 241..244
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 246..251
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 257..263
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 269..275
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 282..284
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 289..302
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 304..318
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 323..326
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 332..334
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 335..339
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 345..357
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 362..365
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 367..370
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 387..390
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 394..403
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 407..412
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 419..421
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 422..426
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 428..430
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 445..448
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 453..467
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 474..477
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 491..509
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 514..517
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 527..530
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 534..537
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 544..554
FT /evidence="ECO:0007829|PDB:2YFO"
FT TURN 555..557
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 560..562
FT /evidence="ECO:0007829|PDB:2YFO"
FT TURN 573..575
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 581..588
FT /evidence="ECO:0007829|PDB:2YFO"
FT TURN 589..592
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 595..597
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 599..601
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 604..626
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 627..631
FT /evidence="ECO:0007829|PDB:2YFO"
FT TURN 635..637
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 638..646
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 650..659
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 680..684
FT /evidence="ECO:0007829|PDB:2YFO"
FT TURN 685..687
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 690..692
FT /evidence="ECO:0007829|PDB:2YFO"
FT HELIX 693..698
FT /evidence="ECO:0007829|PDB:2YFO"
FT STRAND 712..719
FT /evidence="ECO:0007829|PDB:2YFO"
SQ SEQUENCE 935 AA; 105514 MW; BB0D1C0BEAAF0AF6 CRC64;
MAIIYNPNKK IFTLHTAHTT YQMQVDPLGY LLHLYYGEKT NSSMDYVLTY ADRGFSGNPY
AAGMDRTYSL DALPQEYPSL GTGDYRNIAL NIKNEKGVES ADLLFKSYEI RNGKYRLQGL
PAVWADEKEA QTLEIVLADE NAQVEVHLLY GVLEENDVIT RSVRIKNTGT GQITIEKAAA
ACLDFVQGEF DVLRFYGKHA MERNLERTPL GHGTIAFGSR RGTSSHQYNP AVILAEKGTT
ETAGSCYGML FVYSGNFSCE AEKDQFNQTR LLLGLNEELF SYPLASGETF TVPEVILSYS
AEGLSALSQQ YHNCIRNHVC RSKYVHMQRP VLINSWEAAY FDFTGDTIVD LAKEAASLGI
DMVVMDDGWF GKRNDDNSSL GDWQVNETKL GGSLAELITR VHEQGMKFGI WIEPEMINED
SDLYRAHPDW AIRIQGKKPV RSRNQLLLDF SRKEVRDCVF DQICVVLDQG KIDYVKWDMN
RSMADVYAGN LSYDYVLGVY DFMERLCSRY PDLLLEGCSG GGGRFDAGML YYSPQIWCSD
NTDAINRTRI QYGTSFFYPV SAMGAHVSAV PNHQTGRVTS FHTRGVTAMA GTFGYELNPA
LLSDEEKQQI REQIKTYKKY ETLINEGTYW RLSDPFTDEI AAWMSVSEEQ DHALVSVVRL
MAEANQATVY VRLRGLKPDA VYLEEQSGRQ YSGAALMHAG IPLPPFTEEY EAYQFAFTEL
KEAGRLYEKV QKWCDGNAEN RVVISIYGGS GSGKTTLATA LQQYFLNDGT ECYLLSGDDY
PHRIPKRNDE ERMRVYKEAG EDGLRGYLGT KKEIDFDRIN EVLAAFHEGK DSITLRHMGR
EDGEISLEET DFSGISVLLL EWTHGGSDDL HGVDLPVFLE SSPGETRERR IRRNRDENAA
SPFICRVVEL EQEKLEVQRK NAGLIVGKDG SVYEQ
