ADAC_FLAPL
ID ADAC_FLAPL Reviewed; 772 AA.
AC P0DTR4;
DT 13-NOV-2019, integrated into UniProtKB/Swiss-Prot.
DT 13-NOV-2019, sequence version 1.
DT 03-AUG-2022, entry version 10.
DE RecName: Full=A type blood N-acetyl-alpha-D-galactosamine deacetylase {ECO:0000303|PubMed:31182795};
DE EC=3.5.1.- {ECO:0000269|PubMed:31182795};
DE AltName: Full=CBM32 {ECO:0000303|PubMed:31182795};
DE AltName: Full=FpGalNAc deacetylase;
DE Short=FpGalNAcDeAc {ECO:0000303|PubMed:31182795};
DE Flags: Precursor;
OS Flavonifractor plautii (Fusobacterium plautii).
OC Bacteria; Firmicutes; Clostridia; Eubacteriales; Oscillospiraceae;
OC Flavonifractor.
OX NCBI_TaxID=292800;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], X-RAY CRYSTALLOGRAPHY (1.30 ANGSTROMS)
RP OF 28-509 ALONE AND IN COMPLEX WITH SUBSTRATE ANALOG, FUNCTION, CATALYTIC
RP ACTIVITY, REACTION MECHANISM, COFACTOR, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, DOMAIN, BIOTECHNOLOGY, BINDS CARBOHYDRATE,
RP AND MUTAGENESIS OF GLU-90; CYS-125; ASP-126; HIS-278 AND TYR-341.
RX PubMed=31182795; DOI=10.1038/s41564-019-0469-7;
RA Rahfeld P., Sim L., Moon H., Constantinescu I., Morgan-Lang C.,
RA Hallam S.J., Kizhakkedathu J.N., Withers S.G.;
RT "An enzymatic pathway in the human gut microbiome that converts A to
RT universal O type blood.";
RL Nat. Microbiol. 4:1475-1485(2019).
CC -!- FUNCTION: One of an enzyme pair that work together to convert the A
CC antigen to the H antigen of the O blood type, which together release
CC galactosamine. Catalyzes the first step in the conversion, generating
CC the substrate for the subsequent enzyme (FpGalNase, AC P0DTR5). Works
CC on many different A antigen subtypes. Glu-90 probably activates a
CC nucleophilic water molecule to start the deacetylation reaction.
CC {ECO:0000269|PubMed:31182795}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=an N-acetyl-alpha-D-galactosaminyl-(1->3)-[alpha-L-fucosyl-
CC (1->2)]-beta-D-galactosyl derivative + H2O = acetate + an alpha-D-
CC galactosaminyl-(1->3)-[alpha-L-fucosyl-(1->2)]-beta-D-galactosyl
CC derivative; Xref=Rhea:RHEA:14869, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:30089, ChEBI:CHEBI:140559, ChEBI:CHEBI:144802;
CC Evidence={ECO:0000269|PubMed:31182795};
CC -!- COFACTOR:
CC Name=a divalent metal cation; Xref=ChEBI:CHEBI:60240;
CC Evidence={ECO:0000269|PubMed:31182795};
CC -!- ACTIVITY REGULATION: Inhibited by EDTA. {ECO:0000269|PubMed:31182795}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=340 uM for A antigen type 1 penta-MU
CC {ECO:0000269|PubMed:31182795};
CC pH dependence:
CC Optimum pH is 8.0. {ECO:0000269|PubMed:31182795};
CC -!- DOMAIN: The deacetylase domain is in the N-terminus, while the C-
CC terminus has a CBM32-type carbohydrate-binding domain that is not
CC required for activity on soluble substrates. The CBM32 domain binds
CC preferentially to repeating N-acetyl lactosamine structures.
CC {ECO:0000269|PubMed:31182795}.
CC -!- BIOTECHNOLOGY: 5 ug/ml of this enzyme pair converts A blood type to O
CC blood type in an hour, and can be removed by centrifugation, showing
CC the pair can be used for production of universal type donor blood.
CC {ECO:0000269|PubMed:31182795}.
CC -!- MISCELLANEOUS: DNA was isolated from a male human fecal sample of AB+
CC blood type, the sequence was given to UniProtKB by the submitters.
CC {ECO:0000269|PubMed:31182795}.
CC -!- WEB RESOURCE: Name=Protein Spotlight; Note=Dropping barriers - Issue
CC 220 of December 2019;
CC URL="https://web.expasy.org/spotlight/back_issues/220/";
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DR RefSeq; WP_009260926.1; NZ_WKPS01000046.1.
DR PDB; 6N1A; X-ray; 1.60 A; A=28-509.
DR PDB; 6N1B; X-ray; 1.30 A; A=28-509.
DR PDBsum; 6N1A; -.
DR PDBsum; 6N1B; -.
DR AlphaFoldDB; P0DTR4; -.
DR SMR; P0DTR4; -.
DR GO; GO:0016798; F:hydrolase activity, acting on glycosyl bonds; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0008152; P:metabolic process; IEA:UniProtKB-KW.
DR Gene3D; 2.115.10.20; -; 1.
DR InterPro; IPR003343; Big_2.
DR InterPro; IPR000421; FA58C.
DR InterPro; IPR008979; Galactose-bd-like_sf.
DR InterPro; IPR023296; Glyco_hydro_beta-prop_sf.
DR InterPro; IPR008964; Invasin/intimin_cell_adhesion.
DR InterPro; IPR036278; Sialidase_sf.
DR Pfam; PF02368; Big_2; 1.
DR Pfam; PF00754; F5_F8_type_C; 1.
DR SMART; SM00635; BID_2; 1.
DR SUPFAM; SSF49373; SSF49373; 1.
DR SUPFAM; SSF49785; SSF49785; 1.
DR SUPFAM; SSF50939; SSF50939; 1.
DR SUPFAM; SSF75005; SSF75005; 1.
DR PROSITE; PS50022; FA58C_3; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Glycosidase; Hydrolase; Metal-binding; Signal.
FT SIGNAL 1..27
FT /evidence="ECO:0000255"
FT CHAIN 28..772
FT /note="A type blood N-acetyl-alpha-D-galactosamine
FT deacetylase"
FT /id="PRO_0000448571"
FT DOMAIN 494..605
FT /note="F5/8 type C"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00081"
FT REGION 180..402
FT /note="Deacetylase activity"
FT /evidence="ECO:0000269|PubMed:31182795"
FT REGION 502..765
FT /note="CBM32 carbohydrate-binding domain"
FT /evidence="ECO:0000269|PubMed:31182795"
FT REGION 515..772
FT /note="Not required for activity on soluble substrates"
FT /evidence="ECO:0000269|PubMed:31182795"
FT BINDING 87
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:31182795,
FT ECO:0007744|PDB:6N1B"
FT BINDING 123
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:31182795,
FT ECO:0007744|PDB:6N1B"
FT BINDING 126
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT /evidence="ECO:0000269|PubMed:31182795,
FT ECO:0007744|PDB:6N1A, ECO:0007744|PDB:6N1B"
FT BINDING 236
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:31182795,
FT ECO:0007744|PDB:6N1B"
FT BINDING 278
FT /ligand="a divalent metal cation"
FT /ligand_id="ChEBI:CHEBI:60240"
FT /evidence="ECO:0000269|PubMed:31182795,
FT ECO:0007744|PDB:6N1A, ECO:0007744|PDB:6N1B"
FT MUTAGEN 90
FT /note="E->A,L: Loss of deacetylase activity."
FT /evidence="ECO:0000269|PubMed:31182795"
FT MUTAGEN 125
FT /note="C->A,S: 5-fold reduction in deacetylase rate."
FT /evidence="ECO:0000269|PubMed:31182795"
FT MUTAGEN 126
FT /note="D->N: Dramatic reduction in deacetylase rate."
FT /evidence="ECO:0000269|PubMed:31182795"
FT MUTAGEN 278
FT /note="H->A,F: Very dramatic reduction in deacetylase
FT rate."
FT /evidence="ECO:0000269|PubMed:31182795"
FT MUTAGEN 341
FT /note="Y->F: 3000-fold reduction in deacetylase rate."
FT /evidence="ECO:0000269|PubMed:31182795"
SQ SEQUENCE 772 AA; 84441 MW; 55C1AD3F8018D0D7 CRC64;
MRNRRKAVSL LTGLLVTAQL FPTAALAADS SESALNKAPG YQDFPAYYSD SAHADDQVTH
PDVVVLEEPW NGYRYWAVYT PNVMRISIYE NPSIVASSDG VHWVEPEGLS NPIEPQPPST
RYHNCDADMV YNAEYDAMMA YWNWADDQGG GVGAEVRLRI SYDGVHWGVP VTYDEMTRVW
SKPTSDAERQ VADGEDDFIT AIASPDRYDM LSPTIVYDDF RDVFILWANN TGDVGYQNGQ
ANFVEMRYSD DGITWGEPVR VNGFLGLDEN GQQLAPWHQD VQYVPDLKEF VCISQCFAGR
NPDGSVLHLT TSKDGVNWEQ VGTKPLLSPG PDGSWDDFQI YRSSFYYEPG SSAGDGTMRV
WYSALQKDTN NKMVADSSGN LTIQAKSEDD RIWRIGYAEN SFVEMMRVLL DDPGYTTPAL
VSGNSLMLSA ETTSLPTGDV MKLETSFAPV DTSDQVVKYT SSDPDVATVD EFGTITGVSV
GSARIMAETR EGLSDDLEIA VVENPYTLIP QSNMTATATS VYGGTTEGPA SNVLDGNVRT
IWHTNYAPKD ELPQSITVSF DQPYTVGRFV YTPRQNGTNG IISEYELYAI HQDGSKDLVA
SGSDWALDAK DKTVSFAPVE AVGLELKAIA GAGGFGTAAE LNVYAYGPIE PAPVYVPVDD
RDASLVFTGA WNSDSNGSFY EGTARYTNEI GASVEFTFVG TAIRWYGQND VNFGAAEVYV
DGVLAGEVNV YGPAAAQQLL FEADGLAYGK HTIRIVCVSP VVDFDYFSYV GE
