O33830 (AGLA_THEMA) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Alpha-glucosidase
EC=3.2.1.20

Alternative name(s):
Maltase

Gene names

Name:	aglA
Ordered Locus Names:	TM_1834

Organism

Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099) [Reference proteome] [HAMAP]

Taxonomic identifier

243274 [NCBI]

Taxonomic lineage

cellular organisms › Bacteria › Thermotogae › Thermotogae › Thermotogales › Thermotogaceae › Thermotoga › Thermotoga maritima

Protein attributes

Sequence length	480 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Alpha-glycosidase with a very broad specificity. Hydrolyzes maltose and other small maltooligosaccharides but is inactive against the polymeric substrate starch. AglA is not specific with respect to the configuration at the C-4 position of its substrates because glycosidic derivatives of D-galactose are also hydrolyzed. Does not cleave beta-glycosidic bonds.
Catalytic activity	Hydrolysis of terminal, non-reducing (1->4)-linked alpha-D-glucose residues with release of alpha-D-glucose.
Cofactor	Binds 1 NAD per subunit. Ref.3 Binds 1 manganese ion per subunit. Can also use cobalt and nickel ions, albeit less efficiently than manganese ion. Ref.3
Enzyme regulation	Inhibited by Hg2+ ion and EDTA.
Subunit structure	Homodimer. Ref.3
Sequence similarities	Belongs to the glycosyl hydrolase 4 family.
Caution	In the crystal structure (Ref.5), the metal ion is absent, probably due to the oxidation of the active site Cys-174 to sulfinic acid. In the absence of metal, positions of the coenzyme and the substrate and their interactions are all significantly altered, presumably accounting for the inactivity of this form.
Biophysicochemical properties	Kinetic parameters: KM=0.23 mM for p-nitrophenyl-alpha-D-glucopyranoside Ref.3 Ref.4 KM=0.53 mM for p-nitrophenyl-alpha-D-galactopyranoside Vmax=9.94 µmol/min/mg enzyme with p-nitrophenyl-alpha-D-glucopyranoside as substrate Vmax=11.42 µmol/min/mg enzyme with p-nitrophenyl-alpha-D-galactopyranoside as substrate pH dependence: Optimum pH is 7.5. Active from pH 6.5 to 9. Temperature dependence: Optimum temperature is 90 degrees Celsius. Active from 60 to 105 degrees Celsius. Highly thermostable.

Ontologies

Keywords
Biological process	Carbohydrate metabolism
Ligand	Cobalt Manganese Metal-binding NAD Nickel
Molecular function	Glycosidase Hydrolase
Technical term	3D-structure Complete proteome Reference proteome
Gene Ontology (GO)
Biological_process	carbohydrate metabolic process Inferred from electronic annotation. Source: UniProtKB-KW
Molecular_function	maltose alpha-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 – 480

480

Alpha-glucosidase

PRO_0000169855

Regions

Nucleotide binding

4 – 70

67

NAD

Sites

Active site

175

1

Proton donor Probable

Active site

260

1

Proton acceptor Probable

Metal binding

174

1

Manganese By similarity

Metal binding

203

1

Manganese By similarity

Binding site

119

1

Substrate

Binding site

153

1

Substrate

Experimental info

Mutagenesis

10

1

G → A: Reduced activity. 300-fold reduction of the binding affinity for NAD(+). No change in substrate affinity. Ref.4

Mutagenesis

12

1

G → A: No change in activity and substrate affinity. 5-fold reduction of the binding affinity for NAD(+). Ref.4

Mutagenesis

13

1

S → A: Highly reduced activity. 10-fold reduction of the binding affinity for NAD(+). No change in substrate affinity. Ref.4

Sequence conflict

459 – 479

21

Missing Ref.1

Secondary structure

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480

Helix Strand Turn

Details...

Beta strand

5 – 9

5

Turn

10 – 12

3

Helix

14 – 25

12

Helix

28 – 30

3

Beta strand

34 – 38

5

Helix

42 – 58

17

Beta strand

64 – 69

6

Helix

71 – 75

5

Beta strand

79 – 83

5

Helix

89 – 102

14

Beta strand

122 – 124

3

Helix

126 – 142

17

Beta strand

147 – 150

4

Helix

155 – 165

11

Beta strand

168 – 173

6

Helix

176 – 178

3

Helix

179 – 186

8

Helix

191 – 193

3

Beta strand

194 – 201

8

Beta strand

204 – 212

9

Helix

218 – 227

10

Helix

229 – 231

3

Helix

244 – 253

10

Helix

259 – 261

3

Helix

267 – 270

4

Helix

273 – 280

8

Turn

282 – 285

4

Helix

290 – 316

27

Helix

322 – 324

3

Beta strand

326 – 329

4

Helix

333 – 346

14

Helix

356 – 365

10

Beta strand

369 – 376

8

Beta strand

388 – 397

10

Beta strand

400 – 403

4

Helix

412 – 417

6

Helix

419 – 435

17

Helix

438 – 446

9

Helix

454 – 465

12

Helix

468 – 470

3

Helix

471 – 477

7

Sequences

	Sequence		Length	Mass (Da)
	O33830 [UniParc]. Last modified May 30, 2000. Version 2. Checksum: 4CA5DB3B155CE5A1	FASTA	480	55,047
	10 20 30 40 50 60 MPSVKIGIIG AGSAVFSLRL VSDLCKTPGL SGSTVTLMDI DEERLDAILT IAKKYVEEVG 70 80 90 100 110 120 ADLKFEKTMN LDDVIIDADF VINTAMVGGH TYLEKVRQIG EKYGYYRGID AQEFNMVSDY 130 140 150 160 170 180 YTFSNYNQLK YFVDIARKIE KLSPKAWYLQ AANPIFEGTT LVTRTVPIKA VGFCHGHYGV 190 200 210 220 230 240 MEIVEKLGLE EEKVDWQVAG VNHGIWLNRF RYNGGNAYPL LDKWIEEKSK DWKPENPFND 250 260 270 280 290 300 QLSPAAIDMY RFYGVMPIGD TVRNSSWRYH RDLETKKKWY GEPWGGADSE IGWKWYQDTL 310 320 330 340 350 360 GKVTEITKKV AKFIKENPSV RLSDLGSVLG KDLSEKQFVL EVEKILDPER KSGEQHIPFI 370 380 390 400 410 420 DALLNDNKAR FVVNIPNKGI IHGIDDDVVV EVPALVDKNG IHPEKIEPPL PDRVVKYYLR 430 440 450 460 470 480 PRIMRMEMAL EAFLTGDIRI IKELLYRDPR TKSDEQVEKV IEEILALPEN EEMRKHYLKR

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Isolation and analysis of genes for amylolytic enzymes of the hyperthermophilic bacterium Thermotoga maritima." Bibel M., Brettl C., Gosslar U., Kriegshaeuser G., Liebl W. FEMS Microbiol. Lett. 158:9-15(1998) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: ATCC 43589 / MSB8 / DSM 3109 / JCM 10099.
[2]	"Evidence for lateral gene transfer between Archaea and Bacteria from genome sequence of Thermotoga maritima." Nelson K.E., Clayton R.A., Gill S.R., Gwinn M.L., Dodson R.J., Haft D.H., Hickey E.K., Peterson J.D., Nelson W.C., Ketchum K.A., McDonald L.A., Utterback T.R., Malek J.A., Linher K.D., Garrett M.M., Stewart A.M., Cotton M.D., Pratt M.S. , Phillips C.A., Richardson D.L., Heidelberg J.F., Sutton G.G., Fleischmann R.D., Eisen J.A., White O., Salzberg S.L., Smith H.O., Venter J.C., Fraser C.M. Nature 399:323-329(1999) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: ATCC 43589 / MSB8 / DSM 3109 / JCM 10099.
[3]	"Thermotoga maritima AglA, an extremely thermostable NAD+-, Mn2+-, and thiol-dependent alpha-glucosidase." Raasch C., Streit W., Schanzer J., Bibel M., Gosslar U., Liebl W. Extremophiles 4:189-200(2000) [PubMed] [Europe PMC] [Abstract] Cited for: CHARACTERIZATION, SUBUNIT, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES. Strain: ATCC 43589 / MSB8 / DSM 3109 / JCM 10099.
[4]	"Identification of residues important for NAD+ binding by the Thermotoga maritima alpha-glucosidase AglA, a member of glycoside hydrolase family 4." Raasch C., Armbrecht M., Streit W., Hoecker B., Straeter N., Liebl W. FEBS Lett. 517:267-271(2002) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF GLY-10; GLY-12 AND SER-13, KINETIC PARAMETERS.
[5]	"Crystal structure of Thermotoga maritima alpha-glucosidase AglA defines a new clan of NAD+-dependent glycosidases." Lodge J.A., Maier T., Liebl W., Hoffmann V., Straeter N. J. Biol. Chem. 278:19151-19158(2003) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) OF COMPLEX WITH NAD(+) AND MALTOSE.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

AJ001089 Genomic DNA. Translation: CAA04524.1.
AE000512 Genomic DNA. Translation: AAD36897.1.

PIR

F72205.

RefSeq

NP_229631.1. NC_000853.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1OBB	X-ray	1.90	A/B	1-480	[»]

ProteinModelPortal

O33830.

SMR

O33830. Positions 2-479.

ModBase

Search...

Protein-protein interaction databases

STRING

243274.TM1834.

Protein family/group databases

CAZy

GH4. Glycoside Hydrolase Family 4.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

AAD36897; AAD36897; TM_1834.

GeneID

897356.

KEGG

tma:TM1834.

PATRIC

23938655. VBITheMar51294_1854.

Phylogenomic databases

eggNOG

COG1486.

KO

K07406.

OMA

DYHTIWG.

ProtClustDB

CLSK875141.

Enzyme and pathway databases

BRENDA

3.2.1.20. 6331.

Family and domain databases

Gene3D

3.40.50.720. 1 hit.
3.90.110.10. 2 hits.

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

O33830.

Entry information

Entry name

AGLA_THEMA

Accession

Primary (citable) accession number: O33830

Entry history

Integrated into UniProtKB/Swiss-Prot:	December 15, 1998
Last sequence update:	May 30, 2000
Last modified:	May 1, 2013
This is version 105 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program