ID AGLA_THEMA Reviewed; 480 AA. AC O33830; DT 15-DEC-1998, integrated into UniProtKB/Swiss-Prot. DT 30-MAY-2000, sequence version 2. DT 27-MAR-2024, entry version 162. DE RecName: Full=Alpha-glucosidase; DE EC=3.2.1.20; DE AltName: Full=Maltase; GN Name=aglA; OrderedLocusNames=TM_1834; OS Thermotoga maritima (strain ATCC 43589 / DSM 3109 / JCM 10099 / NBRC 100826 OS / MSB8). OC Bacteria; Thermotogota; Thermotogae; Thermotogales; Thermotogaceae; OC Thermotoga. OX NCBI_TaxID=243274; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=ATCC 43589 / DSM 3109 / JCM 10099 / NBRC 100826 / MSB8; RX PubMed=9453151; DOI=10.1111/j.1574-6968.1998.tb12793.x; RA Bibel M., Brettl C., Gosslar U., Kriegshaeuser G., Liebl W.; RT "Isolation and analysis of genes for amylolytic enzymes of the RT hyperthermophilic bacterium Thermotoga maritima."; RL FEMS Microbiol. Lett. 158:9-15(1998). RN [2] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=ATCC 43589 / DSM 3109 / JCM 10099 / NBRC 100826 / MSB8; RX PubMed=10360571; DOI=10.1038/20601; RA Nelson K.E., Clayton R.A., Gill S.R., Gwinn M.L., Dodson R.J., Haft D.H., RA Hickey E.K., Peterson J.D., Nelson W.C., Ketchum K.A., McDonald L.A., RA Utterback T.R., Malek J.A., Linher K.D., Garrett M.M., Stewart A.M., RA Cotton M.D., Pratt M.S., Phillips C.A., Richardson D.L., Heidelberg J.F., RA Sutton G.G., Fleischmann R.D., Eisen J.A., White O., Salzberg S.L., RA Smith H.O., Venter J.C., Fraser C.M.; RT "Evidence for lateral gene transfer between Archaea and Bacteria from RT genome sequence of Thermotoga maritima."; RL Nature 399:323-329(1999). RN [3] RP CHARACTERIZATION, SUBUNIT, COFACTOR, AND BIOPHYSICOCHEMICAL PROPERTIES. RC STRAIN=ATCC 43589 / DSM 3109 / JCM 10099 / NBRC 100826 / MSB8; RX PubMed=10972187; DOI=10.1007/pl00010711; RA Raasch C., Streit W., Schanzer J., Bibel M., Gosslar U., Liebl W.; RT "Thermotoga maritima AglA, an extremely thermostable NAD+-, Mn2+-, and RT thiol-dependent alpha-glucosidase."; RL Extremophiles 4:189-200(2000). RN [4] RP MUTAGENESIS OF GLY-10; GLY-12 AND SER-13, AND KINETIC PARAMETERS. RX PubMed=12062450; DOI=10.1016/s0014-5793(02)02641-8; RA Raasch C., Armbrecht M., Streit W., Hoecker B., Straeter N., Liebl W.; RT "Identification of residues important for NAD+ binding by the Thermotoga RT maritima alpha-glucosidase AglA, a member of glycoside hydrolase family RT 4."; RL FEBS Lett. 517:267-271(2002). RN [5] RP X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) OF COMPLEX WITH NAD(+) AND MALTOSE. RX PubMed=12588867; DOI=10.1074/jbc.m211626200; RA Lodge J.A., Maier T., Liebl W., Hoffmann V., Straeter N.; RT "Crystal structure of Thermotoga maritima alpha-glucosidase AglA defines a RT new clan of NAD+-dependent glycosidases."; RL J. Biol. Chem. 278:19151-19158(2003). CC -!- FUNCTION: Alpha-glycosidase with a very broad specificity. Hydrolyzes CC maltose and other small maltooligosaccharides but is inactive against CC the polymeric substrate starch. AglA is not specific with respect to CC the configuration at the C-4 position of its substrates because CC glycosidic derivatives of D-galactose are also hydrolyzed. Does not CC cleave beta-glycosidic bonds. CC -!- CATALYTIC ACTIVITY: CC Reaction=Hydrolysis of terminal, non-reducing (1->4)-linked alpha-D- CC glucose residues with release of alpha-D-glucose.; EC=3.2.1.20; CC -!- COFACTOR: CC Name=NAD(+); Xref=ChEBI:CHEBI:57540; CC Evidence={ECO:0000269|PubMed:10972187}; CC Note=Binds 1 NAD(+) per subunit. {ECO:0000269|PubMed:10972187}; CC -!- COFACTOR: CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035; CC Evidence={ECO:0000269|PubMed:10972187}; CC Name=Co(2+); Xref=ChEBI:CHEBI:48828; CC Evidence={ECO:0000269|PubMed:10972187}; CC Name=Ni(2+); Xref=ChEBI:CHEBI:49786; CC Evidence={ECO:0000269|PubMed:10972187}; CC Note=Binds 1 Mn(2+) ion per subunit. Can also use Co(2+) and Ni(2+) CC ions, albeit less efficiently than manganese ion. CC {ECO:0000269|PubMed:10972187}; CC -!- ACTIVITY REGULATION: Inhibited by Hg(2+) ion and EDTA. CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=0.23 mM for p-nitrophenyl-alpha-D-glucopyranoside CC {ECO:0000269|PubMed:10972187, ECO:0000269|PubMed:12062450}; CC KM=0.53 mM for p-nitrophenyl-alpha-D-galactopyranoside CC {ECO:0000269|PubMed:10972187, ECO:0000269|PubMed:12062450}; CC Vmax=9.94 umol/min/mg enzyme with CC p-nitrophenyl-alpha-D-glucopyranoside as substrate CC {ECO:0000269|PubMed:10972187, ECO:0000269|PubMed:12062450}; CC Vmax=11.42 umol/min/mg enzyme with CC p-nitrophenyl-alpha-D-galactopyranoside as substrate CC {ECO:0000269|PubMed:10972187, ECO:0000269|PubMed:12062450}; CC pH dependence: CC Optimum pH is 7.5. Active from pH 6.5 to 9. CC {ECO:0000269|PubMed:10972187}; CC Temperature dependence: CC Optimum temperature is 90 degrees Celsius. Active from 60 to 105 CC degrees Celsius. Highly thermostable. {ECO:0000269|PubMed:10972187}; CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:10972187}. CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 4 family. {ECO:0000305}. CC -!- CAUTION: In the crystal structure (PubMed:12588867), the metal ion is CC absent, probably due to the oxidation of the active site Cys-174 to CC sulfinic acid. In the absence of metal, positions of the coenzyme and CC the substrate and their interactions are all significantly altered, CC presumably accounting for the inactivity of this form. CC {ECO:0000305|PubMed:12588867}. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; AJ001089; CAA04524.1; -; Genomic_DNA. DR EMBL; AE000512; AAD36897.1; -; Genomic_DNA. DR PIR; F72205; F72205. DR RefSeq; NP_229631.1; NC_000853.1. DR RefSeq; WP_010865414.1; NZ_CP011107.1. DR PDB; 1OBB; X-ray; 1.90 A; A/B=1-480. DR PDBsum; 1OBB; -. DR AlphaFoldDB; O33830; -. DR SMR; O33830; -. DR STRING; 243274.TM_1834; -. DR DrugBank; DB03323; Maltose. DR CAZy; GH4; Glycoside Hydrolase Family 4. DR PaxDb; 243274-THEMA_05030; -. DR EnsemblBacteria; AAD36897; AAD36897; TM_1834. DR KEGG; tma:TM1834; -. DR eggNOG; COG1486; Bacteria. DR InParanoid; O33830; -. DR OrthoDB; 9808275at2; -. DR BRENDA; 3.2.1.20; 6331. DR SABIO-RK; O33830; -. DR EvolutionaryTrace; O33830; -. DR Proteomes; UP000008183; Chromosome. DR GO; GO:0032450; F:maltose alpha-glucosidase activity; IEA:UniProtKB-EC. DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW. DR GO; GO:0016616; F:oxidoreductase activity, acting on the CH-OH group of donors, NAD or NADP as acceptor; IEA:InterPro. DR CDD; cd05297; GH4_alpha_glucosidase_galactosidase; 1. DR Gene3D; 3.90.1820.10; AglA-like glucosidase; 1. DR InterPro; IPR019802; GlycHydrolase_4_CS. DR InterPro; IPR001088; Glyco_hydro_4. DR InterPro; IPR022616; Glyco_hydro_4_C. DR InterPro; IPR015955; Lactate_DH/Glyco_Ohase_4_C. DR InterPro; IPR036291; NAD(P)-bd_dom_sf. DR NCBIfam; NF041089; alpha_gluc_AglA; 1. DR PANTHER; PTHR32092; 6-PHOSPHO-BETA-GLUCOSIDASE-RELATED; 1. DR PANTHER; PTHR32092:SF4; ALPHA-GLUCOSIDASE; 1. DR Pfam; PF02056; Glyco_hydro_4; 1. DR Pfam; PF11975; Glyco_hydro_4C; 1. DR PRINTS; PR00732; GLHYDRLASE4. DR SUPFAM; SSF56327; LDH C-terminal domain-like; 1. DR SUPFAM; SSF51735; NAD(P)-binding Rossmann-fold domains; 1. DR PROSITE; PS01324; GLYCOSYL_HYDROL_F4; 1. PE 1: Evidence at protein level; KW 3D-structure; Carbohydrate metabolism; Cobalt; Glycosidase; Hydrolase; KW Manganese; Metal-binding; NAD; Nickel; Reference proteome. FT CHAIN 1..480 FT /note="Alpha-glucosidase" FT /id="PRO_0000169855" FT ACT_SITE 175 FT /note="Proton donor" FT /evidence="ECO:0000305" FT ACT_SITE 260 FT /note="Proton acceptor" FT /evidence="ECO:0000305" FT BINDING 4..70 FT /ligand="NAD(+)" FT /ligand_id="ChEBI:CHEBI:57540" FT BINDING 119 FT /ligand="substrate" FT BINDING 153 FT /ligand="substrate" FT BINDING 174 FT /ligand="Mn(2+)" FT /ligand_id="ChEBI:CHEBI:29035" FT /evidence="ECO:0000250" FT BINDING 203 FT /ligand="Mn(2+)" FT /ligand_id="ChEBI:CHEBI:29035" FT /evidence="ECO:0000250" FT MUTAGEN 10 FT /note="G->A: Reduced activity. 300-fold reduction of the FT binding affinity for NAD(+). No change in substrate FT affinity." FT /evidence="ECO:0000269|PubMed:12062450" FT MUTAGEN 12 FT /note="G->A: No change in activity and substrate affinity. FT 5-fold reduction of the binding affinity for NAD(+)." FT /evidence="ECO:0000269|PubMed:12062450" FT MUTAGEN 13 FT /note="S->A: Highly reduced activity. 10-fold reduction of FT the binding affinity for NAD(+). No change in substrate FT affinity." FT /evidence="ECO:0000269|PubMed:12062450" FT CONFLICT 459..479 FT /note="Missing (in Ref. 1)" FT /evidence="ECO:0000305" FT STRAND 5..9 FT /evidence="ECO:0007829|PDB:1OBB" FT TURN 10..12 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 14..25 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 28..30 FT /evidence="ECO:0007829|PDB:1OBB" FT STRAND 34..38 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 42..58 FT /evidence="ECO:0007829|PDB:1OBB" FT STRAND 64..69 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 71..75 FT /evidence="ECO:0007829|PDB:1OBB" FT STRAND 79..83 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 89..102 FT /evidence="ECO:0007829|PDB:1OBB" FT STRAND 122..124 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 126..142 FT /evidence="ECO:0007829|PDB:1OBB" FT STRAND 147..150 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 155..165 FT /evidence="ECO:0007829|PDB:1OBB" FT STRAND 168..173 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 176..178 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 179..186 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 191..193 FT /evidence="ECO:0007829|PDB:1OBB" FT STRAND 194..201 FT /evidence="ECO:0007829|PDB:1OBB" FT STRAND 204..212 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 218..227 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 229..231 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 244..253 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 259..261 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 267..270 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 273..280 FT /evidence="ECO:0007829|PDB:1OBB" FT TURN 282..285 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 290..316 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 322..324 FT /evidence="ECO:0007829|PDB:1OBB" FT STRAND 326..329 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 333..346 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 356..365 FT /evidence="ECO:0007829|PDB:1OBB" FT STRAND 369..376 FT /evidence="ECO:0007829|PDB:1OBB" FT STRAND 388..397 FT /evidence="ECO:0007829|PDB:1OBB" FT STRAND 400..403 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 412..417 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 419..435 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 438..446 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 454..465 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 468..470 FT /evidence="ECO:0007829|PDB:1OBB" FT HELIX 471..477 FT /evidence="ECO:0007829|PDB:1OBB" SQ SEQUENCE 480 AA; 55047 MW; 4CA5DB3B155CE5A1 CRC64; MPSVKIGIIG AGSAVFSLRL VSDLCKTPGL SGSTVTLMDI DEERLDAILT IAKKYVEEVG ADLKFEKTMN LDDVIIDADF VINTAMVGGH TYLEKVRQIG EKYGYYRGID AQEFNMVSDY YTFSNYNQLK YFVDIARKIE KLSPKAWYLQ AANPIFEGTT LVTRTVPIKA VGFCHGHYGV MEIVEKLGLE EEKVDWQVAG VNHGIWLNRF RYNGGNAYPL LDKWIEEKSK DWKPENPFND QLSPAAIDMY RFYGVMPIGD TVRNSSWRYH RDLETKKKWY GEPWGGADSE IGWKWYQDTL GKVTEITKKV AKFIKENPSV RLSDLGSVLG KDLSEKQFVL EVEKILDPER KSGEQHIPFI DALLNDNKAR FVVNIPNKGI IHGIDDDVVV EVPALVDKNG IHPEKIEPPL PDRVVKYYLR PRIMRMEMAL EAFLTGDIRI IKELLYRDPR TKSDEQVEKV IEEILALPEN EEMRKHYLKR //