AMT4_PSEST
ID AMT4_PSEST Reviewed; 548 AA.
AC P13507;
DT 01-JAN-1990, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1997, sequence version 2.
DT 03-AUG-2022, entry version 136.
DE RecName: Full=Glucan 1,4-alpha-maltotetraohydrolase;
DE Short=G4-amylase;
DE EC=3.2.1.60 {ECO:0000269|PubMed:10556241, ECO:0000269|PubMed:2646279, ECO:0000269|PubMed:9281429};
DE AltName: Full=Exo-maltotetraohydrolase;
DE AltName: Full=Maltotetraose-forming amylase;
DE AltName: Full=Maltotetraose-forming exo-amylase;
DE Flags: Precursor;
GN Name=amyP;
OS Pseudomonas stutzeri (Pseudomonas perfectomarina).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=316;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PARTIAL PROTEIN SEQUENCE, AND CATALYTIC
RP ACTIVITY.
RC STRAIN=MO-19;
RX PubMed=2646279; DOI=10.1128/jb.171.3.1333-1339.1989;
RA Fujita M., Torigoe K., Nakada T., Tsusaki K., Kubota M., Sakai S.,
RA Tsujisaka Y.;
RT "Cloning and nucleotide sequence of the gene (amyP) for maltotetraose-
RT forming amylase from Pseudomonas stutzeri MO-19.";
RL J. Bacteriol. 171:1333-1339(1989).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 22-439 IN COMPLEX WITH CALCIUM,
RP SEQUENCE REVISION TO 286-302, DISULFIDE BONDS, AND COFACTOR.
RC STRAIN=MO-19;
RX PubMed=9126844; DOI=10.1006/jmbi.1996.0887;
RA Morishita Y., Hasegawa K., Matsuura Y., Katsube Y., Kubota M., Sakai S.;
RT "Crystal structure of a maltotetraose-forming exo-amylase from Pseudomonas
RT stutzeri.";
RL J. Mol. Biol. 267:661-672(1997).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) OF MUTANT GLN-240 IN COMPLEX WITH
RP MALTOPENTAOSE AND CALCIUM IONS, DISULFIDE BONDS, MUTAGENESIS OF GLU-240,
RP CATALYTIC ACTIVITY, AND COFACTOR.
RC STRAIN=MO-19;
RX PubMed=9281429; DOI=10.1006/jmbi.1997.1222;
RA Yoshioka Y., Hasegawa K., Matsuura Y., Katsube Y., Kubota M.;
RT "Crystal structures of a mutant maltotetraose-forming exo-amylase
RT cocrystallized with maltopentaose.";
RL J. Mol. Biol. 271:619-628(1997).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 22-450 IN COMPLEX WITH CALCIUM
RP AND MALTOTETRAOSE, DISULFIDE BONDS, ACTIVE SITE, MUTAGENESIS OF ASP-214;
RP GLU-240 AND ASP-315, CATALYTIC ACTIVITY, AND COFACTOR.
RC STRAIN=MO-19;
RX PubMed=10556241; DOI=10.1093/protein/12.10.819;
RA Hasegawa K., Kubota M., Matsuura Y.;
RT "Roles of catalytic residues in alpha-amylases as evidenced by the
RT structures of the product-complexed mutants of a maltotetraose-forming
RT amylase.";
RL Protein Eng. 12:819-824(1999).
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Hydrolysis of (1->4)-alpha-D-glucosidic linkages in amylaceous
CC polysaccharides, to remove successive maltotetraose residues from the
CC non-reducing chain ends.; EC=3.2.1.60;
CC Evidence={ECO:0000269|PubMed:10556241, ECO:0000269|PubMed:2646279,
CC ECO:0000269|PubMed:9281429};
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000269|PubMed:10556241, ECO:0000269|PubMed:9126844,
CC ECO:0000269|PubMed:9281429};
CC Note=Binds 2 calcium ions per subunit. {ECO:0000269|PubMed:10556241,
CC ECO:0000269|PubMed:9126844, ECO:0000269|PubMed:9281429};
CC -!- PATHWAY: Glycan degradation; starch degradation.
CC {ECO:0000305|PubMed:10556241, ECO:0000305|PubMed:9281429}.
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:9281429}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000305|PubMed:2646279}.
CC -!- MISCELLANEOUS: There are several isoenzyme forms of this protein.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 13 family. {ECO:0000305}.
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DR EMBL; M24516; AAA25707.1; -; Genomic_DNA.
DR PIR; A32803; A32803.
DR PDB; 1GCY; X-ray; 1.60 A; A=22-548.
DR PDB; 1JDA; X-ray; 2.20 A; A=22-450.
DR PDB; 1JDC; X-ray; 1.90 A; A=22-450.
DR PDB; 1JDD; X-ray; 1.90 A; A=22-450.
DR PDB; 1QI3; X-ray; 2.00 A; A=22-450.
DR PDB; 1QI4; X-ray; 2.00 A; A=22-450.
DR PDB; 1QI5; X-ray; 2.00 A; A=22-450.
DR PDB; 1QPK; X-ray; 2.00 A; A=22-439.
DR PDB; 2AMG; X-ray; 2.00 A; A=22-439.
DR PDBsum; 1GCY; -.
DR PDBsum; 1JDA; -.
DR PDBsum; 1JDC; -.
DR PDBsum; 1JDD; -.
DR PDBsum; 1QI3; -.
DR PDBsum; 1QI4; -.
DR PDBsum; 1QI5; -.
DR PDBsum; 1QPK; -.
DR PDBsum; 2AMG; -.
DR AlphaFoldDB; P13507; -.
DR SMR; P13507; -.
DR STRING; 32042.PstZobell_12666; -.
DR DrugBank; DB02379; Beta-D-Glucose.
DR DrugBank; DB02237; Maltotetraose.
DR CAZy; CBM20; Carbohydrate-Binding Module Family 20.
DR CAZy; GH13; Glycoside Hydrolase Family 13.
DR UniPathway; UPA00153; -.
DR EvolutionaryTrace; P13507; -.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0004556; F:alpha-amylase activity; IEA:InterPro.
DR GO; GO:0033910; F:glucan 1,4-alpha-maltotetraohydrolase activity; IEA:UniProtKB-EC.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:2001070; F:starch binding; IEA:InterPro.
DR GO; GO:0005983; P:starch catabolic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 2.60.40.10; -; 1.
DR Gene3D; 2.60.40.1180; -; 1.
DR InterPro; IPR006046; Alpha_amylase.
DR InterPro; IPR015165; AMT4_domain_C.
DR InterPro; IPR013784; Carb-bd-like_fold.
DR InterPro; IPR002044; CBM_fam20.
DR InterPro; IPR006047; Glyco_hydro_13_cat_dom.
DR InterPro; IPR013780; Glyco_hydro_b.
DR InterPro; IPR017853; Glycoside_hydrolase_SF.
DR InterPro; IPR013783; Ig-like_fold.
DR Pfam; PF00128; Alpha-amylase; 1.
DR Pfam; PF00686; CBM_20; 1.
DR Pfam; PF09081; DUF1921; 1.
DR PRINTS; PR00110; ALPHAAMYLASE.
DR SMART; SM00642; Aamy; 1.
DR SMART; SM01065; CBM_2; 1.
DR SUPFAM; SSF49452; SSF49452; 1.
DR SUPFAM; SSF51445; SSF51445; 1.
DR PROSITE; PS51166; CBM20; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Calcium; Carbohydrate metabolism; Direct protein sequencing;
KW Disulfide bond; Glycosidase; Hydrolase; Metal-binding; Secreted; Signal.
FT SIGNAL 1..21
FT CHAIN 22..548
FT /note="Glucan 1,4-alpha-maltotetraohydrolase"
FT /id="PRO_0000001420"
FT DOMAIN 446..548
FT /note="CBM20"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00594"
FT REGION 529..548
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 214
FT /note="Nucleophile"
FT /evidence="ECO:0000305|PubMed:10556241"
FT ACT_SITE 240
FT /note="Proton donor"
FT /evidence="ECO:0000305|PubMed:10556241,
FT ECO:0000305|PubMed:9281429"
FT BINDING 22
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:10556241,
FT ECO:0000269|PubMed:9126844, ECO:0000269|PubMed:9281429"
FT BINDING 23
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:10556241,
FT ECO:0000269|PubMed:9126844, ECO:0000269|PubMed:9281429"
FT BINDING 34
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:10556241,
FT ECO:0000269|PubMed:9126844, ECO:0000269|PubMed:9281429"
FT BINDING 37
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:10556241,
FT ECO:0000269|PubMed:9126844, ECO:0000269|PubMed:9281429"
FT BINDING 38
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:10556241,
FT ECO:0000269|PubMed:9126844, ECO:0000269|PubMed:9281429"
FT BINDING 99..100
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:10556241"
FT BINDING 137
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:10556241,
FT ECO:0000269|PubMed:9126844, ECO:0000269|PubMed:9281429"
FT BINDING 138
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:10556241,
FT ECO:0000305|PubMed:9281429"
FT BINDING 172
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:10556241,
FT ECO:0000269|PubMed:9126844, ECO:0000269|PubMed:9281429"
FT BINDING 175
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:10556241,
FT ECO:0000269|PubMed:9126844, ECO:0000269|PubMed:9281429"
FT BINDING 177..181
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:10556241"
FT BINDING 183
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:10556241,
FT ECO:0000269|PubMed:9126844, ECO:0000269|PubMed:9281429"
FT BINDING 212
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:10556241,
FT ECO:0000305|PubMed:9281429"
FT BINDING 217..218
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P0C1B3"
FT BINDING 218
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:10556241,
FT ECO:0000269|PubMed:9126844, ECO:0000269|PubMed:9281429"
FT BINDING 314
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:10556241,
FT ECO:0000305|PubMed:9281429"
FT BINDING 326
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:10556241"
FT SITE 315
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000305|PubMed:10556241"
FT DISULFID 161..171
FT /evidence="ECO:0000269|PubMed:10556241,
FT ECO:0000269|PubMed:9126844, ECO:0000269|PubMed:9281429"
FT DISULFID 237..272
FT /evidence="ECO:0000269|PubMed:10556241,
FT ECO:0000269|PubMed:9126844, ECO:0000269|PubMed:9281429"
FT MUTAGEN 214
FT /note="D->N: Abolishes enzyme activity."
FT /evidence="ECO:0000269|PubMed:10556241"
FT MUTAGEN 240
FT /note="E->Q: Abolishes enzyme activity."
FT /evidence="ECO:0000269|PubMed:10556241,
FT ECO:0000269|PubMed:9281429"
FT MUTAGEN 315
FT /note="D->N: Abolishes enzyme activity."
FT /evidence="ECO:0000269|PubMed:10556241"
FT CONFLICT 286..302
FT /note="GSIADWKHGLNGNPDPR -> ARSPTGSTPERQSRPA (in Ref. 1; AA
FT sequence)"
FT /evidence="ECO:0000305"
FT HELIX 34..36
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 40..42
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 48..51
FT /evidence="ECO:0007829|PDB:1GCY"
FT TURN 53..55
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 56..69
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 73..77
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 87..89
FT /evidence="ECO:0007829|PDB:1JDC"
FT STRAND 92..94
FT /evidence="ECO:0007829|PDB:1JDC"
FT STRAND 102..104
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 108..110
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 113..125
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 129..134
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 152..155
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 158..160
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 165..167
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 172..174
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 180..184
FT /evidence="ECO:0007829|PDB:1QI5"
FT HELIX 189..205
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 208..214
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 216..218
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 221..231
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 235..239
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 245..247
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 253..256
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 259..270
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 277..285
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 288..293
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 295..297
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 301..304
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 307..309
FT /evidence="ECO:0007829|PDB:1GCY"
FT TURN 314..316
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 322..324
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 333..335
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 336..345
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 346..353
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 354..358
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 363..376
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 383..386
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 390..398
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 403..409
FT /evidence="ECO:0007829|PDB:1GCY"
FT HELIX 415..417
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 424..429
FT /evidence="ECO:0007829|PDB:1GCY"
FT TURN 430..433
FT /evidence="ECO:0007829|PDB:1GCY"
FT STRAND 434..438
FT /evidence="ECO:0007829|PDB:1GCY"
SQ SEQUENCE 548 AA; 59876 MW; 2B87217B3379158F CRC64;
MSHILRAAVL AAMLLPLPSM ADQAGKSPNA VRYHGGDEII LQGFHWNVVR EAPNDWYNIL
RQQAATIAAD GFSAIWMPVP WRDFSSWSDG SKSGGGEGYF WHDFNKNGRY GSDAQLRQAA
SALGGAGVKV LYDVVPNHMN RGYPDKEINL PAGQGFWRND CADPGNYPND CDDGDRFIGG
DADLNTGHPQ VYGMFRDEFT NLRSQYGAGG FRFDFVRGYA PERVNSWMTD SADNSFCVGE
LWKGPSEYPN WDWRNTASWQ QIIKDWSDRA KCPVFDFALK ERMQNGSIAD WKHGLNGNPD
PRWREVAVTF VDNHDTGYSP GQNGGQHHWA LQDGLIRQAY AYILTSPGTP VVYWSHMYDW
GYGDFIRQLI QVRRAAGVRA DSAISFHSGY SGLVATVSGS QQTLVVALNS DLGNPGQVAS
GSFSEAVNAS NGQVRVWRSG TGSGGGEPGA LVSVSFRCDN GATQMGDSVY AVGNVSQLGN
WSPAAALRLT DTSGYPTWKG SIALPAGQNE EWKCLIRNEA NATQVRQWQG GANNSLTPSE
GATTVGRL
