H2018_AKKM8
ID H2018_AKKM8 Reviewed; 493 AA.
AC B2UP57;
DT 02-JUN-2021, integrated into UniProtKB/Swiss-Prot.
DT 01-JUL-2008, sequence version 1.
DT 03-AUG-2022, entry version 74.
DE RecName: Full=Beta-hexosaminidase Amuc_2018 {ECO:0000305};
DE EC=3.2.1.52 {ECO:0000269|PubMed:29304441, ECO:0000269|PubMed:30846208};
DE AltName: Full=Beta-N-acetylhexosaminidase Am2301 {ECO:0000303|PubMed:29304441, ECO:0000303|PubMed:30846208};
DE Flags: Precursor;
GN OrderedLocusNames=Amuc_2018 {ECO:0000312|EMBL:ACD05828.1};
OS Akkermansia muciniphila (strain ATCC BAA-835 / DSM 22959 / JCM 33894 / BCRC
OS 81048 / CCUG 64013 / CIP 107961 / Muc).
OC Bacteria; Verrucomicrobia; Verrucomicrobiae; Verrucomicrobiales;
OC Akkermansiaceae; Akkermansia.
OX NCBI_TaxID=349741;
RN [1] {ECO:0000312|Proteomes:UP000001031}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC BAA-835 / DSM 22959 / JCM 33894 / BCRC 81048 / CCUG 64013 / CIP
RC 107961 / Muc;
RX PubMed=21390229; DOI=10.1371/journal.pone.0016876;
RA van Passel M.W., Kant R., Zoetendal E.G., Plugge C.M., Derrien M.,
RA Malfatti S.A., Chain P.S., Woyke T., Palva A., de Vos W.M., Smidt H.;
RT "The genome of Akkermansia muciniphila, a dedicated intestinal mucin
RT degrader, and its use in exploring intestinal metagenomes.";
RL PLoS ONE 6:E16876-E16876(2011).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND SUBSTRATE SPECIFICITY.
RC STRAIN=ATCC BAA-835 / DSM 22959 / JCM 33894 / BCRC 81048 / CCUG 64013 / CIP
RC 107961 / Muc {ECO:0000303|PubMed:29304441};
RX PubMed=29304441; DOI=10.1016/j.carres.2017.12.007;
RA Wang M., Zhang X.Y., Guo R.R., Cai Z.P., Hu X.C., Chen H., Wei S.,
RA Voglmeir J., Liu L.;
RT "Cloning, purification and biochemical characterization of two beta-N-
RT acetylhexosaminidases from the mucin-degrading gut bacterium Akkermansia
RT muciniphila.";
RL Carbohydr. Res. 457:1-7(2018).
RN [3] {ECO:0007744|PDB:6JE8, ECO:0007744|PDB:6JEA, ECO:0007744|PDB:6JEB}
RP X-RAY CRYSTALLOGRAPHY (1.50 ANGSTROMS) OF 22-490 IN COMPLEXES WITH
RP N-ACETYLGLUCOSAMINE AND ZINC, FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, ACTIVE SITES, AND MUTAGENESIS OF ASP-278;
RP GLU-279 AND TYR-373.
RX PubMed=30846208; DOI=10.1016/j.bbrc.2019.02.074;
RA Chen X., Wang J., Liu M., Yang W., Wang Y., Tang R., Zhang M.;
RT "Crystallographic evidence for substrate-assisted catalysis of beta-N-
RT acetylhexosaminidas from Akkermansia muciniphila.";
RL Biochem. Biophys. Res. Commun. 511:833-839(2019).
CC -!- FUNCTION: Hydrolyzes terminal GlcNAc residues from terminally
CC unbranched N-glycans and from chitobiose. Hydrolyzes beta-1,6-linked N-
CC acetylglucosamine and beta-1,4-linked N-acetylgalactosamine from pNP-
CC alpha-GalNAc[beta1,3Gal]beta1,6GlcNAc and pNP-beta-GlcNAc-beta1,4-
CC GalNAc substrates, respectively, as well as beta-1,2-linked N-
CC acetylglucosamine units from the non-reducing end of N-glycans.
CC Hydrolyzes GlcNAc residues linked to alpha1,3- or alpha1,6-mannose
CC branch, but has low activity on substrates with more than one GlcNAc
CC residue on one of the mannose branches. Releases terminal GlcNAc
CC moieties from the N-glycopeptide Gly-Glu-Asn-(GlcNAc2Man3GlcNAc2)-Arg
CC with high efficiency. Has moderate hydrolytic activity on the
CC chitobiose moiety of N-glycopeptide substrate Gly-Glu-Asn-(GlcNAc2)-
CC Arg. Does not hydrolyze GlcNAc residues from N-glycan structures
CC bearing a bisecting GlcNAc moiety (beta1,4-linked GlcNAc to the
CC beta1,4-linked core mannose) (PubMed:29304441). Potentially capable of
CC cleaving the specific glycoside linkages in the process of mucin
CC degradation in human intestinal tract (Probable). Hydrolyzes synthetic
CC substrate pNP-beta-GlcNAc with high activity and pNP-beta-GalNAc to a
CC lesser extent (PubMed:29304441, PubMed:30846208). Does not hydrolyze
CC pNP-beta-glucose, pNP-beta-galactose, pNP-alpha-glucose, pNP-alpha-
CC galactose, pNP-alpha-GlcNAc or pNP-alpha-fucose (PubMed:29304441).
CC {ECO:0000269|PubMed:29304441, ECO:0000269|PubMed:30846208,
CC ECO:0000305}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Hydrolysis of terminal non-reducing N-acetyl-D-hexosamine
CC residues in N-acetyl-beta-D-hexosaminides.; EC=3.2.1.52;
CC Evidence={ECO:0000269|PubMed:29304441, ECO:0000269|PubMed:30846208};
CC -!- ACTIVITY REGULATION: Significantly inhibited by the addition of sodium
CC dodecyl sulfate (SDS), but not by EDTA, urea, 2-mercaptoethanol or
CC Triton X-100. Strongly inhibited by Cu2(+) ions, in case of which the
CC activity is decreased by 70%. No significant inhibition with Al(3+),
CC Fe(3+), Ca(2+), Cd(2+), Mg(2+), Mn(2+), Ni(2+) and Zn(2+) ions.
CC Strongly inhibited by PugNAc (O-(2-acetamido-2-deoxy-D-
CC glucopyranosylideneamino) N-phenylcarbamate) in the sub-micromolar
CC concentration range. PugNAc at a concentration of 0.5 mM decreases the
CC activity by 50% and the addition of 1 mM PugNAc fully inhibits the
CC enzyme. No significant reduction in the activity by alkylation using N-
CC ethylmaleimide or 2-iodoacetamide. {ECO:0000269|PubMed:29304441}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.031 mM for pNP-beta-GlcNAc (at pH 7.0 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:30846208};
CC KM=0.52 mM for pNP-beta-GlcNAc (at pH 5.0 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:29304441};
CC KM=0.11 mM for pNP-beta-GalNAc (at pH 5.0 and 37 degrees Celsius)
CC {ECO:0000269|PubMed:29304441};
CC Vmax=79.99 umol/min/mg enzyme with pNP-beta-GlcNAc as substrate (at
CC pH 7.0 and 37 degrees Celsius) {ECO:0000269|PubMed:30846208};
CC Vmax=44.0 umol/min/mg enzyme with pNP-beta-GlcNAc as substrate (at pH
CC 5.0 and 37 degrees Celsius) {ECO:0000269|PubMed:29304441};
CC Note=kcat is 1.01 sec(-1) and kcat/KM is 34.34 mM(-1)sec(-1) with
CC pNP-beta-GlcNAc as substrate. {ECO:0000269|PubMed:30846208};
CC pH dependence:
CC Optimum pH is 5.0. The activity drops sharply below pH 4.5 and above
CC pH 5.5. {ECO:0000269|PubMed:29304441};
CC Temperature dependence:
CC Optimum temperature is 37 degrees Celsius. Retains more than 90% of
CC activity after 96 hours of incubation at 37 degrees Celsius and after
CC 12 hours of incubation at 42 degrees Celsius.
CC {ECO:0000269|PubMed:29304441};
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 20 family. {ECO:0000305}.
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DR EMBL; CP001071; ACD05828.1; -; Genomic_DNA.
DR RefSeq; WP_012421042.1; NC_010655.1.
DR PDB; 6JE8; X-ray; 1.80 A; A=22-490.
DR PDB; 6JEA; X-ray; 2.27 A; A=22-490.
DR PDB; 6JEB; X-ray; 1.50 A; A=22-490.
DR PDBsum; 6JE8; -.
DR PDBsum; 6JEA; -.
DR PDBsum; 6JEB; -.
DR AlphaFoldDB; B2UP57; -.
DR SMR; B2UP57; -.
DR STRING; 349741.Amuc_2018; -.
DR CAZy; GH20; Glycoside Hydrolase Family 20.
DR EnsemblBacteria; ACD05828; ACD05828; Amuc_2018.
DR KEGG; amu:Amuc_2018; -.
DR eggNOG; COG3525; Bacteria.
DR HOGENOM; CLU_007082_5_1_0; -.
DR OrthoDB; 727559at2; -.
DR BioCyc; AMUC349741:G1GBX-2149-MON; -.
DR Proteomes; UP000001031; Chromosome.
DR GO; GO:0032428; F:beta-N-acetylgalactosaminidase activity; IDA:UniProtKB.
DR GO; GO:0016231; F:beta-N-acetylglucosaminidase activity; IDA:UniProtKB.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0102148; F:N-acetyl-beta-D-galactosaminidase activity; IEA:UniProtKB-EC.
DR GO; GO:0052781; P:chitobiose catabolic process; IDA:UniProtKB.
DR GO; GO:0000272; P:polysaccharide catabolic process; IDA:UniProtKB.
DR Gene3D; 3.30.379.10; -; 1.
DR InterPro; IPR025705; Beta_hexosaminidase_sua/sub.
DR InterPro; IPR015883; Glyco_hydro_20_cat.
DR InterPro; IPR017853; Glycoside_hydrolase_SF.
DR InterPro; IPR029018; Hex-like_dom2.
DR InterPro; IPR015882; HEX_bac_N.
DR PANTHER; PTHR22600; PTHR22600; 1.
DR Pfam; PF00728; Glyco_hydro_20; 1.
DR Pfam; PF02838; Glyco_hydro_20b; 1.
DR PIRSF; PIRSF001093; B-hxosamndse_ab_euk; 1.
DR PRINTS; PR00738; GLHYDRLASE20.
DR SUPFAM; SSF51445; SSF51445; 1.
DR SUPFAM; SSF55545; SSF55545; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Carbohydrate metabolism; Glycosidase; Hydrolase;
KW Metal-binding; Polysaccharide degradation; Reference proteome; Signal;
KW Zinc.
FT SIGNAL 1..21
FT /evidence="ECO:0000255"
FT CHAIN 22..493
FT /note="Beta-hexosaminidase Amuc_2018"
FT /evidence="ECO:0000255"
FT /id="PRO_0000452893"
FT ACT_SITE 151
FT /note="Charge relay system"
FT /evidence="ECO:0000305|PubMed:30846208,
FT ECO:0007744|PDB:6JE8, ECO:0007744|PDB:6JEA,
FT ECO:0007744|PDB:6JEB"
FT ACT_SITE 214
FT /note="Charge relay system"
FT /evidence="ECO:0000305|PubMed:30846208,
FT ECO:0007744|PDB:6JE8, ECO:0007744|PDB:6JEA,
FT ECO:0007744|PDB:6JEB"
FT ACT_SITE 279
FT /note="Charge relay system"
FT /evidence="ECO:0000269|PubMed:30846208,
FT ECO:0007744|PDB:6JEB"
FT BINDING 122
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:30846208,
FT ECO:0007744|PDB:6JEA"
FT BINDING 227
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000269|PubMed:30846208,
FT ECO:0007744|PDB:6JE8, ECO:0007744|PDB:6JEA,
FT ECO:0007744|PDB:6JEB"
FT BINDING 247
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000269|PubMed:30846208,
FT ECO:0007744|PDB:6JE8, ECO:0007744|PDB:6JEA,
FT ECO:0007744|PDB:6JEB"
FT BINDING 278
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:30846208,
FT ECO:0007744|PDB:6JEA"
FT BINDING 288
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000269|PubMed:30846208,
FT ECO:0007744|PDB:6JE8, ECO:0007744|PDB:6JEA,
FT ECO:0007744|PDB:6JEB"
FT BINDING 291
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000269|PubMed:30846208,
FT ECO:0007744|PDB:6JE8, ECO:0007744|PDB:6JEA,
FT ECO:0007744|PDB:6JEB"
FT BINDING 345
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:30846208,
FT ECO:0007744|PDB:6JEA"
FT BINDING 373..375
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:30846208,
FT ECO:0007744|PDB:6JEA"
FT BINDING 421..423
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:30846208,
FT ECO:0007744|PDB:6JEA"
FT MUTAGEN 278
FT /note="D->A: No significant change in KM value, but 37-fold
FT and 2600-fold decrease in specific activity and kcat/KM
FT value compared to that of wild-type, respectively."
FT /evidence="ECO:0000269|PubMed:30846208"
FT MUTAGEN 279
FT /note="E->A: No significant change in KM value, but 8.7-
FT fold and 3500-fold decrease in specific activity and
FT kcat/KM value compared to that of wild-type, respectively."
FT /evidence="ECO:0000269|PubMed:30846208"
FT MUTAGEN 373
FT /note="Y->F: No significant change in KM value, but 6.3-
FT fold and 1226-fold decrease in specific activity and
FT kcat/KM value compared to that of wild-type, respectively."
FT /evidence="ECO:0000269|PubMed:30846208"
FT STRAND 32..42
FT /evidence="ECO:0007829|PDB:6JEB"
FT STRAND 46..51
FT /evidence="ECO:0007829|PDB:6JEB"
FT STRAND 60..65
FT /evidence="ECO:0007829|PDB:6JEB"
FT STRAND 68..75
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 76..94
FT /evidence="ECO:0007829|PDB:6JEB"
FT STRAND 100..107
FT /evidence="ECO:0007829|PDB:6JEB"
FT STRAND 112..118
FT /evidence="ECO:0007829|PDB:6JEB"
FT TURN 120..122
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 127..139
FT /evidence="ECO:0007829|PDB:6JEB"
FT STRAND 144..148
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 164..167
FT /evidence="ECO:0007829|PDB:6JEB"
FT TURN 168..170
FT /evidence="ECO:0007829|PDB:6JEB"
FT STRAND 171..173
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 188..199
FT /evidence="ECO:0007829|PDB:6JEB"
FT TURN 200..202
FT /evidence="ECO:0007829|PDB:6JEB"
FT STRAND 204..208
FT /evidence="ECO:0007829|PDB:6JEB"
FT STRAND 212..214
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 216..221
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 223..225
FT /evidence="ECO:0007829|PDB:6JEB"
FT STRAND 226..228
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 251..267
FT /evidence="ECO:0007829|PDB:6JEB"
FT STRAND 272..274
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 283..287
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 289..298
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 303..320
FT /evidence="ECO:0007829|PDB:6JEB"
FT STRAND 324..328
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 333..335
FT /evidence="ECO:0007829|PDB:6JEB"
FT STRAND 341..344
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 350..360
FT /evidence="ECO:0007829|PDB:6JEB"
FT STRAND 363..366
FT /evidence="ECO:0007829|PDB:6JEB"
FT TURN 369..371
FT /evidence="ECO:0007829|PDB:6JEB"
FT TURN 385..387
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 393..397
FT /evidence="ECO:0007829|PDB:6JEB"
FT TURN 401..404
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 407..410
FT /evidence="ECO:0007829|PDB:6JEB"
FT STRAND 413..420
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 428..435
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 438..447
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 450..452
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 455..473
FT /evidence="ECO:0007829|PDB:6JEB"
FT HELIX 481..483
FT /evidence="ECO:0007829|PDB:6JEB"
SQ SEQUENCE 493 AA; 56044 MW; 5EC0C54715B8946C CRC64;
MARPLPILGG ILLSFSPPAE ATAQYSIIPE PSRTELRQET AKTLQLLSDQ EVPTLETDAY
RLTVTPQGAH LASGGREGRI YGLATLRQLR DQLAGQPEGI PCGVITDKPR YPWRGLMVDP
ARHFIPAADL KKFVDMMAYY KFNRLHLHLT DNQGWRLPVP GYPKLKSVAS RREESFGDGI
PHEGMYTKQE LKELVAYCAA RGIDVIPEID MPGHNQALHA AYPEFFCFPK PDMNVRTTAG
NSKELVCPQK PEVWKFYASV FNELKDIFPS GIVHLGGDEA PTELWEKCPL CREARTRAAM
KDEQEQMKAF FAKTAALLAK NGQTPQFWYE GNAGIYHPGE TVYAWRQGQA LQSIEKTKKA
GLNLIMASSE YCYLDFPQIQ GQRNWGWMKT TTLQKCYDLD PAFGKPEKEA GHIRGVHAPV
WAERLPDLNH LLYRAYPRAC AIAEAGWSPM GVRSWENFRR KLADHRQFIL KRFNYDMERT
QGNEPAFRWE NNK
