ISOA_PSEAY
ID ISOA_PSEAY Reviewed; 776 AA.
AC P10342;
DT 01-JUL-1989, integrated into UniProtKB/Swiss-Prot.
DT 15-DEC-1998, sequence version 3.
DT 03-AUG-2022, entry version 124.
DE RecName: Full=Isoamylase;
DE EC=3.2.1.68 {ECO:0000269|PubMed:2248978};
DE Flags: Precursor;
GN Name=iam;
OS Pseudomonas amyloderamosa.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales.
OX NCBI_TaxID=32043;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=SB-15;
RX PubMed=3379068; DOI=10.1016/s0021-9258(19)76535-1;
RA Amemura A., Chakraborty R., Fujita M., Noumi T., Futai M.;
RT "Cloning and nucleotide sequence of the isoamylase gene from Pseudomonas
RT amyloderamosa SB-15.";
RL J. Biol. Chem. 263:9271-9275(1988).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], CATALYTIC ACTIVITY, AND SUBCELLULAR
RP LOCATION.
RC STRAIN=JD210;
RX PubMed=2248978; DOI=10.1016/0167-4781(90)90004-l;
RA Chen J.H., Chen Z.Y., Chow T.Y., Chen J.C., Tan S.T., Hsu W.H.;
RT "Nucleotide sequence and expression of the isoamylase gene from an
RT isoamylase-hyperproducing mutant, Pseudomonas amyloderamosa JD210.";
RL Biochim. Biophys. Acta 1087:309-315(1990).
RN [3]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 744-776.
RC STRAIN=SB-15;
RX PubMed=2753857; DOI=10.1128/jb.171.8.4320-4325.1989;
RA Amemura A., Fujita M., Futai M.;
RT "Transcription of the isoamylase gene (iam) in Pseudomonas amyloderamosa
RT SB-15.";
RL J. Bacteriol. 171:4320-4325(1989).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 27-776 IN COMPLEX WITH CALCIUM,
RP DISULFIDE BONDS, AND COFACTOR.
RX PubMed=9719642; DOI=10.1006/jmbi.1998.1992;
RA Katsuya Y., Mezaki Y., Kubota M., Matsuura Y.;
RT "Three-dimensional structure of Pseudomonas isoamylase at 2.2-A
RT resolution.";
RL J. Mol. Biol. 281:885-897(1998).
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Hydrolysis of (1->6)-alpha-D-glucosidic branch linkages in
CC glycogen, amylopectin and their beta-limit dextrins.; EC=3.2.1.68;
CC Evidence={ECO:0000269|PubMed:2248978};
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000269|PubMed:9719642};
CC Note=Binds 1 Ca(2+) ion per subunit. {ECO:0000269|PubMed:9719642};
CC -!- SUBUNIT: Monomer.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:2248978}.
CC -!- INDUCTION: By maltose.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 13 family. {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=AAA25854.1; Type=Frameshift; Evidence={ECO:0000305};
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DR EMBL; J03871; AAA25854.1; ALT_FRAME; Genomic_DNA.
DR EMBL; X13378; CAA31754.1; -; Genomic_DNA.
DR PDB; 1BF2; X-ray; 2.00 A; A=27-776.
DR PDBsum; 1BF2; -.
DR AlphaFoldDB; P10342; -.
DR SMR; P10342; -.
DR CAZy; CBM48; Carbohydrate-Binding Module Family 48.
DR CAZy; GH13; Glycoside Hydrolase Family 13.
DR PRIDE; P10342; -.
DR KEGG; ag:AAA25854; -.
DR BRENDA; 3.2.1.68; 5091.
DR EvolutionaryTrace; P10342; -.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0004133; F:glycogen debranching enzyme activity; IEA:UniProt.
DR GO; GO:0019156; F:isoamylase activity; IEA:UniProtKB-EC.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR CDD; cd02856; E_set_GDE_Isoamylase_N; 1.
DR Gene3D; 2.60.40.10; -; 1.
DR Gene3D; 2.60.40.1180; -; 1.
DR InterPro; IPR044505; GlgX_Isoamylase_N_E_set.
DR InterPro; IPR006047; Glyco_hydro_13_cat_dom.
DR InterPro; IPR004193; Glyco_hydro_13_N.
DR InterPro; IPR013780; Glyco_hydro_b.
DR InterPro; IPR017853; Glycoside_hydrolase_SF.
DR InterPro; IPR013783; Ig-like_fold.
DR InterPro; IPR014756; Ig_E-set.
DR Pfam; PF00128; Alpha-amylase; 1.
DR Pfam; PF02922; CBM_48; 1.
DR SMART; SM00642; Aamy; 1.
DR SUPFAM; SSF51445; SSF51445; 1.
DR SUPFAM; SSF81296; SSF81296; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Calcium; Disulfide bond; Glycosidase; Hydrolase;
KW Metal-binding; Secreted; Signal.
FT SIGNAL 1..26
FT CHAIN 27..776
FT /note="Isoamylase"
FT /id="PRO_0000001424"
FT ACT_SITE 401
FT /note="Nucleophile"
FT ACT_SITE 461
FT /note="Proton donor"
FT BINDING 154
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:9719642,
FT ECO:0007744|PDB:1BF2"
FT BINDING 255
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:9719642,
FT ECO:0007744|PDB:1BF2"
FT BINDING 256
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:9719642,
FT ECO:0007744|PDB:1BF2"
FT BINDING 258
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:9719642,
FT ECO:0007744|PDB:1BF2"
FT BINDING 285
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:9719642,
FT ECO:0007744|PDB:1BF2"
FT SITE 536
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000250"
FT DISULFID 410..422
FT /evidence="ECO:0000269|PubMed:9719642,
FT ECO:0007744|PDB:1BF2"
FT DISULFID 546..616
FT /evidence="ECO:0000269|PubMed:9719642,
FT ECO:0007744|PDB:1BF2"
FT DISULFID 738..766
FT /evidence="ECO:0000269|PubMed:9719642,
FT ECO:0007744|PDB:1BF2"
FT CONFLICT 8
FT /note="A -> G (in Ref. 1; AAA25854)"
FT /evidence="ECO:0000305"
FT CONFLICT 126
FT /note="F -> C (in Ref. 1; AAA25854)"
FT /evidence="ECO:0000305"
FT CONFLICT 169
FT /note="G -> C (in Ref. 1; AAA25854)"
FT /evidence="ECO:0000305"
FT CONFLICT 386
FT /note="L -> V (in Ref. 1; AAA25854)"
FT /evidence="ECO:0000305"
FT CONFLICT 413..416
FT /note="GAYT -> AVH (in Ref. 1; AAA25854)"
FT /evidence="ECO:0000305"
FT CONFLICT 555..556
FT /note="WP -> S (in Ref. 1; AAA25854)"
FT /evidence="ECO:0000305"
FT CONFLICT 658..661
FT /note="LRPS -> SPV (in Ref. 1; AAA25854)"
FT /evidence="ECO:0000305"
FT STRAND 34..37
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 41..49
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 54..66
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 70..74
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 82..88
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 89..94
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 102..109
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 163..166
FT /evidence="ECO:0007829|PDB:1BF2"
FT TURN 170..174
FT /evidence="ECO:0007829|PDB:1BF2"
FT TURN 178..180
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 202..204
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 207..210
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 212..216
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 224..226
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 230..235
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 237..243
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 247..251
FT /evidence="ECO:0007829|PDB:1BF2"
FT TURN 259..262
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 286..288
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 296..310
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 314..319
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 328..333
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 343..351
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 358..361
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 364..368
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 376..391
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 397..400
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 403..407
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 411..414
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 437..444
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 454..460
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 480..482
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 484..495
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 504..511
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 515..518
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 519..521
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 525..527
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 528..530
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 535..537
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 540..543
FT /evidence="ECO:0007829|PDB:1BF2"
FT TURN 572..577
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 581..597
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 598..605
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 608..610
FT /evidence="ECO:0007829|PDB:1BF2"
FT TURN 626..628
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 636..654
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 656..658
FT /evidence="ECO:0007829|PDB:1BF2"
FT TURN 666..668
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 669..672
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 676..678
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 681..684
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 691..696
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 698..701
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 707..712
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 714..716
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 718..721
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 726..736
FT /evidence="ECO:0007829|PDB:1BF2"
FT HELIX 739..741
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 754..758
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 762..765
FT /evidence="ECO:0007829|PDB:1BF2"
FT STRAND 767..776
FT /evidence="ECO:0007829|PDB:1BF2"
SQ SEQUENCE 776 AA; 83627 MW; F738BF8040246169 CRC64;
MKCPKILAAL LGCAVLAGVP AMPAHAAINS MSLGASYDAQ QANITFRVYS SQATRIVLYL
YSAGYGVQES ATYTLSPAGS GVWAVTVPVS SIKAAGITGA VYYGYRAWGP NWPYASNWGK
GSQAGFVSDV DANGDRFNPN KLLLDPYAQE VSQDPLNPSN QNGNVFASGA SYRTTDSGIY
APKGVVLVPS TQSTGTKPTR AQKDDVIYEV HVRGFTEQDT SIPAQYRGTY YGAGLKASYL
ASLGVTAVEF LPVQETQNDA NDVVPNSDAN QNYWGYMTEN YFSPDRRYAY NKAAGGPTAE
FQAMVQAFHN AGIKVYMDVV YNHTAEGGTW TSSDPTTATI YSWRGLDNAT YYELTSGNQY
FYDNTGIGAN FNTYNTVAQN LIVDSLAYWA NTMGVDGFRF DLASVLGNSC LNGAYTASAP
NCPNGGYNFD AADSNVAINR ILREFTVRPA AGGSGLDLFA EPWAIGGNSY QLGGFPQGWS
EWNGLFRDSL RQAQNELGSM TIYVTQDAND FSGSSNLFQS SGRSPWNSIN FIDVHDGMTL
KDVYSCNGAN NSQAWPYGPS DGGTSTNYSW DQGMSAGTGA AVDQRRAART GMAFEMLSAG
TPLMQGGDEY LRTLQCNNNA YNLDSSANWL TYSWTTDQSN FYTFAQRLIA FRKAHPALRP
SSWYSGSQLT WYQPSGAVAD SNYWNNTSNY AIAYAINGPS LGDSNSIYVA YNGWSSSVTF
TLPAPPSGTQ WYRVTDTCDW NDGASTFVAP GSETLIGGAG TTYGQCGQSL LLLISK
