P69327 (AMYG_ASPAW) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Glucoamylase
EC=3.2.1.3

Alternative name(s):
1,4-alpha-D-glucan glucohydrolase
Glucan 1,4-alpha-glucosidase

Gene names

Name:

GLAA

Organism

Aspergillus awamori (Black koji mold)

Taxonomic identifier

105351 [NCBI]

Taxonomic lineage

cellular organisms › Eukaryota › Opisthokonta › Fungi › Dikarya › Ascomycota › saccharomyceta › Pezizomycotina › leotiomyceta › Eurotiomycetes › Eurotiomycetidae › Eurotiales › Trichocomaceae › mitosporic Trichocomaceae › Aspergillus

Protein attributes

Sequence length	640 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is further processed into a mature form.
Protein existence	Evidence at protein level

General annotation (Comments)

Catalytic activity	Hydrolysis of terminal (1->4)-linked alpha-D-glucose residues successively from non-reducing ends of the chains with release of beta-D-glucose.
Sequence similarities	Belongs to the glycosyl hydrolase 15 family. Contains 1 CBM20 (carbohydrate binding type-20) domain.

Ontologies

Keywords
Biological process	Carbohydrate metabolism Polysaccharide degradation
Coding sequence diversity	Alternative splicing
Domain	Signal
Molecular function	Glycosidase Hydrolase
PTM	Cleavage on pair of basic residues Disulfide bond Glycoprotein
Technical term	3D-structure
Gene Ontology (GO)
Biological_process	polysaccharide catabolic process Inferred from electronic annotation. Source: UniProtKB-KW
Molecular_function	glucan 1,4-alpha-glucosidase activity Inferred from electronic annotation. Source: EC starch binding Inferred from electronic annotation. Source: InterPro
Complete GO annotation...

Alternative products

This entry describes 2 isoforms produced by alternative splicing. [Align] [Select]

Isoform G1 (identifier: P69327-1)

This isoform has been chosen as the 'canonical' sequence. All positional information in this entry refers to it. This is also the sequence that appears in the downloadable versions of the entry.

Isoform G2 (identifier: P69327-2)

The sequence of this isoform differs from the canonical sequence as follows:
     527-534: STSSTSCT → TTRSGMSL
     535-640: Missing.

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Signal peptide

1 – 18

18

Potential

Propeptide

19 – 24

6

PRO_0000001459

Chain

25 – 640

616

Glucoamylase

PRO_0000001460

Regions

Domain

533 – 640

108

CBM20

Sites

Active site

200

1

Proton acceptor Ref.3

Active site

203

1

Proton donor Ref.3

Binding site

144

1

Substrate

Amino acid modifications

Glycosylation

195

1

N-linked (GlcNAc...)

CAR_000112

Glycosylation

419

1

N-linked (GlcNAc...)

CAR_000113

Glycosylation

465

1

O-linked (Man) By similarity

Glycosylation

467

1

O-linked (Man)

Glycosylation

468

1

O-linked (Man)

Glycosylation

476

1

O-linked (Man)

Glycosylation

477

1

O-linked (Man)

Glycosylation

483

1

O-linked (Man)

Glycosylation

484

1

O-linked (Man)

Glycosylation

486

1

O-linked (Man)

Glycosylation

488

1

O-linked (Man)

Glycosylation

492

1

O-linked (Man) By similarity

Glycosylation

496

1

O-linked (Man) By similarity

Glycosylation

499

1

O-linked (Man) By similarity

Glycosylation

500

1

O-linked (Man) By similarity

Glycosylation

501

1

O-linked (Man) By similarity

Glycosylation

502

1

O-linked (Man) By similarity

Glycosylation

504

1

O-linked (Man) By similarity

Glycosylation

506

1

O-linked (Man) By similarity

Glycosylation

508

1

O-linked (Man) By similarity

Glycosylation

510

1

O-linked (Man) By similarity

Glycosylation

512

1

O-linked (Man) By similarity

Glycosylation

513

1

O-linked (Man) By similarity

Glycosylation

514

1

O-linked (Man) By similarity

Glycosylation

515

1

O-linked (Man) By similarity

Glycosylation

517

1

O-linked (Man) By similarity

Glycosylation

518

1

O-linked (Man) By similarity

Glycosylation

520

1

O-linked (Man) By similarity

Glycosylation

522

1

O-linked (Man) By similarity

Glycosylation

524

1

O-linked (Man) By similarity

Glycosylation

525

1

O-linked (Man) By similarity

Glycosylation

526

1

O-linked (Man) By similarity

Glycosylation

527

1

O-linked (Man) By similarity

Glycosylation

528

1

O-linked (Man) By similarity

Glycosylation

529

1

O-linked (Man) By similarity

Glycosylation

530

1

O-linked (Man) By similarity

Glycosylation

531

1

O-linked (Man) By similarity

Glycosylation

532

1

O-linked (Man) By similarity

Glycosylation

534

1

O-linked (Man) By similarity

Glycosylation

535

1

O-linked (Man) By similarity

Disulfide bond

234 ↔ 237

Ref.6

Disulfide bond

246 ↔ 473

Ref.6

Disulfide bond

286 ↔ 294

Ref.6

Natural variations

Alternative sequence

527 – 534

8

STSSTSCT → TTRSGMSL in isoform G2.

VSP_012836

Alternative sequence

535 – 640

106

Missing in isoform G2.

VSP_000262

Experimental info

Mutagenesis

144

1

W → Y: 2% of wild-type activity. Ref.4

Secondary structure

1

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640

Helix Strand Turn

Details...

Helix

26 – 44

19

Turn

47 – 49

3

Beta strand

67 – 71

5

Beta strand

74 – 76

3

Helix

77 – 92

16

Helix

96 – 98

3

Helix

99 – 114

16

Turn

122 – 125

4

Helix

127 – 129

3

Helix

150 – 168

19

Helix

172 – 177

6

Helix

179 – 193

15

Beta strand

206 – 209

4

Helix

210 – 229

20

Helix

235 – 248

14

Helix

249 – 251

3

Beta strand

254 – 257

4

Beta strand

259 – 262

4

Helix

271 – 277

7

Turn

288 – 291

4

Helix

296 – 307

12

Turn

308 – 312

5

Helix

314 – 316

3

Helix

335 – 337

3

Helix

342 – 362

21

Beta strand

364 – 367

4

Turn

369 – 371

3

Helix

372 – 378

7

Beta strand

384 – 388

5

Helix

392 – 415

24

Beta strand

424 – 426

3

Turn

428 – 430

3

Beta strand

433 – 438

6

Helix

440 – 453

14

Helix

463 – 465

3

Sequences

	Sequence		Length	Mass (Da)
	Isoform G1 [UniParc]. Last modified November 1, 1986. Version 1. Checksum: 26F58DD18AD7F702	FASTA	640	68,309
	10 20 30 40 50 60 MSFRSLLALS GLVCTGLANV ISKRATLDSW LSNEATVART AILNNIGADG AWVSGADSGI 70 80 90 100 110 120 VVASPSTDNP DYFYTWTRDS GLVLKTLVDL FRNGDTSLLS TIENYISAQA IVQGISNPSG 130 140 150 160 170 180 DLSSGAGLGE PKFNVDETAY TGSWGRPQRD GPALRATAMI GFGQWLLDNG YTSTATDIVW 190 200 210 220 230 240 PLVRNDLSYV AQYWNQTGYD LWEEVNGSSF FTIAVQHRAL VEGSAFATAV GSSCSWCDSQ 250 260 270 280 290 300 APEILCYLQS FWTGSFILAN FDSSRSGKDA NTLLGSIHTF DPEAACDDST FQPCSPRALA 310 320 330 340 350 360 NHKEVVDSFR SIYTLNDGLS DSEAVAVGRY PEDTYYNGNP WFLCTLAAAE QLYDALYQWD 370 380 390 400 410 420 KQGSLEVTDV SLDFFKALYS DAATGTYSSS SSTYSSIVDA VKTFADGFVS IVETHAASNG 430 440 450 460 470 480 SMSEQYDKSD GEQLSARDLT WSYAALLTAN NRRNSVVPAS WGETSASSVP GTCAATSAIG 490 500 510 520 530 540 TYSSVTVTSW PSIVATGGTT TTATPTGSGS VTSTSKTTAT ASKTSTSTSS TSCTTPTAVA 550 560 570 580 590 600 VTFDLTATTT YGENIYLVGS ISQLGDWETS DGIALSADKY TSSDPLWYVT VTLPAGESFE 610 620 630 640 YKFIRIESDD SVEWESDPNR EYTVPQACGT STATVTDTWR
	Isoform G2 [UniParc]. Checksum: 94D188E946D0C12E	FASTA	534	56,730
	10 20 30 40 50 60 MSFRSLLALS GLVCTGLANV ISKRATLDSW LSNEATVART AILNNIGADG AWVSGADSGI 70 80 90 100 110 120 VVASPSTDNP DYFYTWTRDS GLVLKTLVDL FRNGDTSLLS TIENYISAQA IVQGISNPSG 130 140 150 160 170 180 DLSSGAGLGE PKFNVDETAY TGSWGRPQRD GPALRATAMI GFGQWLLDNG YTSTATDIVW 190 200 210 220 230 240 PLVRNDLSYV AQYWNQTGYD LWEEVNGSSF FTIAVQHRAL VEGSAFATAV GSSCSWCDSQ 250 260 270 280 290 300 APEILCYLQS FWTGSFILAN FDSSRSGKDA NTLLGSIHTF DPEAACDDST FQPCSPRALA 310 320 330 340 350 360 NHKEVVDSFR SIYTLNDGLS DSEAVAVGRY PEDTYYNGNP WFLCTLAAAE QLYDALYQWD 370 380 390 400 410 420 KQGSLEVTDV SLDFFKALYS DAATGTYSSS SSTYSSIVDA VKTFADGFVS IVETHAASNG 430 440 450 460 470 480 SMSEQYDKSD GEQLSARDLT WSYAALLTAN NRRNSVVPAS WGETSASSVP GTCAATSAIG 490 500 510 520 530 TYSSVTVTSW PSIVATGGTT TTATPTGSGS VTSTSKTTAT ASKTSTTTRS GMSL

References

[1]	"Molecular cloning and characterization of the glucoamylase gene of Aspergillus awamori." Nunberg J.H., Meade J.H., Cole G., Lawyer F.C., McCabe P., Schweickart V., Tal R., Wittman V.P., Flatgaard J.E., Innis M.A. Mol. Cell. Biol. 4:2306-2315(1984) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA] (ISOFORMS G1 AND G2).
[2]	Nunberg J.H., Meade J.H., Cole G., Lawyer F.C., McCabe P., Schweickart V., Tal R., Wittman V.P., Flatgaard J.E., Innis M.A. Submitted (FEB-1985) to the EMBL/GenBank/DDBJ databases Cited for: SEQUENCE REVISION.
[3]	"Catalytic mechanism of fungal glucoamylase as defined by mutagenesis of Asp176, Glu179 and Glu180 in the enzyme from Aspergillus awamori." Sierks M.R., Ford C., Reilly P.J., Svensson B. Protein Eng. 3:193-198(1990) [PubMed] [Europe PMC] [Abstract] Cited for: ACTIVE SITES, MUTAGENESIS.
[4]	"Site-directed mutagenesis at the active site Trp120 of Aspergillus awamori glucoamylase." Sierks M.R., Ford C., Reilly P.J., Svensson B. Protein Eng. 2:621-625(1989) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS OF TRP-144.
[5]	"Functional roles and subsite locations of Leu177, Trp178 and Asn182 of Aspergillus awamori glucoamylase determined by site-directed mutagenesis." Sierks M.R., Ford C., Reilly P.J., Svensson B. Protein Eng. 6:75-79(1993) [PubMed] [Europe PMC] [Abstract] Cited for: MUTAGENESIS.
[6]	"Crystal structure of glucoamylase from Aspergillus awamori var. X100 to 2.2-A resolution." Aleshin A., Golubev A., Firsov L.M., Honzatko R.B. J. Biol. Chem. 267:19291-19298(1992) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF 25-495, GLYCOSYLATION, DISULFIDE BONDS. Strain: Var. X100.
[7]	"Refined structure for the complex of acarbose with glucoamylase from Aspergillus awamori var. X100 to 2.4-A resolution." Aleshin A., Firsov L.M., Honzatko R.B. J. Biol. Chem. 269:15631-15639(1994) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS) OF 25-495. Strain: Var. X100.
[8]	"Refined crystal structures of glucoamylase from Aspergillus awamori var. X100." Aleshin A., Hoffman C., Firsov L.M., Honzatko R.B. J. Mol. Biol. 238:575-591(1994) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF 25-495, GLYCOSYLATION. Strain: Var. X100.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

K02465 mRNA. Translation: AAB59296.1.
K02465 mRNA. Translation: AAB59297.1.

PIR

A29166.
A29776. A93066.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1AGM	X-ray	2.30	A	25-495	[»]
1DOG	X-ray	2.30	A	25-495	[»]
1GAH	X-ray	2.00	A	25-496	[»]
1GAI	X-ray	1.70	A	25-497	[»]
1GLM	X-ray	2.40	A	25-495	[»]
3GLY	X-ray	2.20	A	25-495	[»]

ProteinModelPortal

P69327.

SMR

P69327. Positions 25-640.

ModBase

Search...

Protein family/group databases

mycoCLAP

GLA15A_ASPAW.

PTM databases

GlycoSuiteDB

P04064.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Family and domain databases

Gene3D

1.50.10.10. 1 hit.
2.60.40.10. 1 hit.

InterPro

IPR008928. 6-hairpin_glycosidase-like.
IPR012341. 6hp_glycosidase.
IPR013784. Carb-bd-like_fold.
IPR002044. CBM_fam20.
IPR000165. Glucoamylase.
IPR008291. Glucoamylase_SBD.
IPR015902. Glyco_hydro_13.
IPR011613. Glyco_hydro_15.
IPR013783. Ig-like_fold.
[Graphical view]

PANTHER

PTHR10357. PTHR10357. 1 hit.

Pfam

PF00686. CBM_20. 1 hit.
PF00723. Glyco_hydro_15. 1 hit.
[Graphical view]

PIRSF

PIRSF001031. Glu-a-glcsd_SBD. 1 hit.

PRINTS

PR00736. GLHYDRLASE15.

SMART

SM01065. CBM_2. 1 hit.
[Graphical view]

SUPFAM

SSF49452. CBD_4. 1 hit.
SSF48208. Glyco_trans_6hp. 1 hit.

PROSITE

PS51166. CBM20. 1 hit.
PS00820. GLUCOAMYLASE. 1 hit.
[Graphical view]

ProtoNet

Search...

Other

BindingDB

P69327.

EvolutionaryTrace

P69327.

Entry information

Entry name

AMYG_ASPAW

Accession

Primary (citable) accession number: P69327
Secondary accession number(s): P04064, Q92201, Q99179

Entry history

Integrated into UniProtKB/Swiss-Prot:	November 1, 1986
Last sequence update:	November 1, 1986
Last modified:	April 3, 2013
This is version 55 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Fungal Protein Annotation Program