HEXC_OSTFU
ID HEXC_OSTFU Reviewed; 594 AA.
AC Q06GJ0;
DT 07-APR-2021, integrated into UniProtKB/Swiss-Prot.
DT 31-OCT-2006, sequence version 1.
DT 25-MAY-2022, entry version 93.
DE RecName: Full=Chitooligosaccharidolytic beta-N-acetylglucosaminidase {ECO:0000305};
DE EC=3.2.1.52 {ECO:0000269|PubMed:18959754, ECO:0000269|PubMed:21106526, ECO:0000269|PubMed:21692744, ECO:0000269|PubMed:23300622, ECO:0000269|PubMed:25155420, ECO:0000269|PubMed:25436416, ECO:0000269|PubMed:30135205};
DE AltName: Full=Beta-GlcNAcase {ECO:0000303|PubMed:25155420, ECO:0000303|PubMed:25436416};
DE AltName: Full=Beta-N-acetyl-D-hexosaminidase {ECO:0000303|PubMed:18959754, ECO:0000303|PubMed:21106526, ECO:0000303|PubMed:21692744, ECO:0000303|PubMed:23300622, ECO:0000303|PubMed:30135205};
DE AltName: Full=Beta-hexosaminidase {ECO:0000255|PIRNR:PIRNR001093};
DE AltName: Full=OfHex1 {ECO:0000303|PubMed:18959754, ECO:0000303|PubMed:21106526, ECO:0000303|PubMed:21692744, ECO:0000303|PubMed:23300622, ECO:0000303|PubMed:25155420, ECO:0000303|PubMed:25436416, ECO:0000303|PubMed:30135205};
DE Flags: Precursor;
OS Ostrinia furnacalis (Asian corn borer).
OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota;
OC Neoptera; Endopterygota; Lepidoptera; Glossata; Ditrysia; Pyraloidea;
OC Crambidae; Pyraustinae; Ostrinia.
OX NCBI_TaxID=93504 {ECO:0000312|EMBL:ABI81756.1};
RN [1] {ECO:0000312|EMBL:ABI81756.1}
RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 108-118; 110-118; 128-141;
RP 142-154; 177-199; 200-212; 213-220; 260-265 AND 500-513, FUNCTION,
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE SPECIFICITY,
RP REACTION MECHANISM, SUBUNIT, DEVELOPMENTAL STAGE, IDENTIFICATION BY MASS
RP SPECTROMETRY, MASS SPECTROMETRY, AND BIOTECHNOLOGY.
RX PubMed=18959754; DOI=10.1111/j.1742-4658.2008.06695.x;
RA Yang Q., Liu T., Liu F., Qu M., Qian X.;
RT "A novel beta-N-acetyl-D-hexosaminidase from the insect Ostrinia furnacalis
RT (Guenee).";
RL FEBS J. 275:5690-5702(2008).
RN [2] {ECO:0007744|PDB:3OZP, ECO:0007744|PDB:3S6T}
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 23-594 AND MUTANT GLY-327 IN
RP COMPLEXES WITH PUGNAC INHIBITOR, CATALYTIC ACTIVITY, ACTIVITY REGULATION,
RP SUBUNIT, DISULFIDE BONDS, AND MUTAGENESIS OF VAL-327.
RX PubMed=21692744; DOI=10.1042/bj20110390;
RA Liu T., Zhang H., Liu F., Chen L., Shen X., Yang Q.;
RT "Active-pocket size differentiating insectile from bacterial chitinolytic
RT beta-N-acetyl-D-hexosaminidases.";
RL Biochem. J. 438:467-474(2011).
RN [3] {ECO:0007744|PDB:3NSM, ECO:0007744|PDB:3NSN}
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF 23-594 AND IN COMPLEX WITH
RP TMG-CHITOTRIOMYCIN INHIBITOR, FUNCTION, CATALYTIC ACTIVITY, ACTIVITY
RP REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, REACTION MECHANISM, SUBUNIT,
RP DEVELOPMENTAL STAGE, DISRUPTION PHENOTYPE, DISULFIDE BONDS, AND MUTAGENESIS
RP OF VAL-327; GLU-328; HIS-433; TRP-448 AND TRP-490.
RX PubMed=21106526; DOI=10.1074/jbc.m110.184796;
RA Liu T., Zhang H., Liu F., Wu Q., Shen X., Yang Q.;
RT "Structural determinants of an insect beta-N-Acetyl-D-hexosaminidase
RT specialized as a chitinolytic enzyme.";
RL J. Biol. Chem. 286:4049-4058(2011).
RN [4] {ECO:0007744|PDB:3VTR}
RP X-RAY CRYSTALLOGRAPHY (2.50 ANGSTROMS) OF 23-594 OF MUTANT ALA-328 IN
RP COMPLEX WITH TMG-CHITOTRIOMYCIN INHIBITOR, CATALYTIC ACTIVITY, ACTIVITY
RP REGULATION, SITES, DISULFIDE BONDS, AND MUTAGENESIS OF VAL-327; GLU-328 AND
RP TRP-490.
RX PubMed=23300622; DOI=10.1371/journal.pone.0052225;
RA Liu T., Zhou Y., Chen L., Chen W., Liu L., Shen X., Zhang W., Zhang J.,
RA Yang Q.;
RT "Structural insights into cellulolytic and chitinolytic enzymes revealing
RT crucial residues of insect beta-N-acetyl-D-hexosaminidase.";
RL PLoS ONE 7:e52225-e52225(2012).
RN [5] {ECO:0007744|PDB:3WMB, ECO:0007744|PDB:3WMC}
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF 23-594 IN COMPLEXES WITH
RP NAPHTHALIMIDE DERIVATIVE INHIBITORS, CATALYTIC ACTIVITY, ACTIVITY
RP REGULATION, BIOTECHNOLOGY, GLYCOSYLATION AT ASN-164 AND ASN-375, DISULFIDE
RP BONDS, AND MUTAGENESIS OF TRP-490.
RX PubMed=25155420; DOI=10.1038/srep06188;
RA Liu T., Guo P., Zhou Y., Wang J., Chen L., Yang H., Qian X., Yang Q.;
RT "A crystal structure-guided rational design switching non-carbohydrate
RT inhibitors' specificity between two beta-GlcNAcase homologs.";
RL Sci. Rep. 4:6188-6188(2014).
RN [6] {ECO:0007744|PDB:3OZO}
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 23-594 IN COMPLEX WITH
RP NAG-THIAZOLINE INHIBITOR, CATALYTIC ACTIVITY, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, REACTION MECHANISM, DISULFIDE BONDS, AND
RP MUTAGENESIS OF GLU-328.
RX PubMed=25436416; DOI=10.1016/j.febslet.2014.11.032;
RA Liu T., Xia M., Zhang H., Zhou H., Wang J., Shen X., Yang Q.;
RT "Exploring NAG-thiazoline and its derivatives as inhibitors of chitinolytic
RT beta-acetylglucosaminidases.";
RL FEBS Lett. 589:110-116(2015).
RN [7] {ECO:0007744|PDB:5Y0V, ECO:0007744|PDB:5Y1B}
RP X-RAY CRYSTALLOGRAPHY (2.21 ANGSTROMS) OF 23-594 IN COMPLEXES WITH
RP BERBERINE INHIBITOR AND ITS DERIVATIVE, CATALYTIC ACTIVITY, ACTIVITY
RP REGULATION, GLYCOSYLATION AT ASN-164 AND ASN-375, DISULFIDE BONDS, AND
RP MUTAGENESIS OF TRP-490.
RX PubMed=30135205; DOI=10.1074/jbc.ra118.004351;
RA Duan Y., Liu T., Zhou Y., Dou T., Yang Q.;
RT "Glycoside hydrolase family 18 and 20 enzymes are novel targets of the
RT traditional medicine berberine.";
RL J. Biol. Chem. 293:15429-15438(2018).
CC -!- FUNCTION: Hydrolyzes one beta-GlcNAc unit at a time from the non-
CC reducing ends of substrates, with a preference for shorter substrates.
CC The 2-acetamido group and the beta-glycoside bond linkage in the
CC substrate are required for its activity. Active with p-nitrophenyl
CC (pNP)-beta-GlcNAc, pNP-beta-GalNAc and chitooligosaccharides (degree of
CC polymerization from 2 to 6), but not with the complex N-glycan
CC substrate (GlcNAcbeta-1,2Manalpha-1,6)(GlcNAcbeta-1,2Manalpha-
CC 1,3)Manbeta-1,4GlcNAcbeta-1,4GlcNAc-PA (GnGn-PA), pNP-alpha-GlcNAc or
CC with the long polymer colloidal chitin. Involved in chitin catabolism
CC (PubMed:18959754). Involved in the degradation of old cuticle during
CC the pupation stage (PubMed:21106526). {ECO:0000269|PubMed:18959754,
CC ECO:0000269|PubMed:21106526}.
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:0000255|PIRNR:PIRNR001093, ECO:0000269|PubMed:18959754,
CC ECO:0000269|PubMed:21106526, ECO:0000269|PubMed:21692744,
CC ECO:0000269|PubMed:23300622, ECO:0000269|PubMed:25155420,
CC ECO:0000269|PubMed:25436416, ECO:0000269|PubMed:30135205};
CC -!- ACTIVITY REGULATION: Inhibited by O-(2-acetamido-2-deoxy-D-
CC glucopyransylidene)-amino-N-phenylcarbamate (PUGNAc) (PubMed:21692744).
CC Inhibited by thiabendazole (TMG)-chitotriomycin (PubMed:21692744,
CC PubMed:21106526, PubMed:23300622). Inhibited by 6-(dimethylamino)-2-(2-
CC (((5-methyl-1,3,4-thiadiazol-2-yl)methyl)amino)ethyl)- 1H-
CC benzo[de]isoquinoline-1,3(2H)-dione (Q2), a synthesized non-
CC carbohydrate unsymmetrical dyad of naphthalimide and thiadiazole having
CC a dimethylamino group at C4 of the naphthalimide (PubMed:25155420).
CC Inhibited poorly by N-acetyl-glucosamine (NAG)-thiazoline (NGT), but
CC when the thiazoline ring of NGT is replaced by a bulky substituent such
CC as in compound 1,2-dideoxy-2'-methylamino-alpha-D-glucopyranoso-[2,1-
CC d]-Delta2'-thiazoline (NMAGT), the inhibition constant Ki is lowered
CC 600-fold compared to that of NGT (PubMed:25436416). Inhibited by
CC berberine, berberine analogs thalifendine and palmatine, and berberine
CC derivative SYSU-1, but not by berberine analog tetrahydroberberine
CC (PubMed:30135205). {ECO:0000269|PubMed:21106526,
CC ECO:0000269|PubMed:21692744, ECO:0000269|PubMed:23300622,
CC ECO:0000269|PubMed:25155420, ECO:0000269|PubMed:25436416,
CC ECO:0000269|PubMed:30135205}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.30 mM for GlcNAc-beta-1,4-GlcNAc (at pH 6.8 and 25 degrees
CC Celsius) {ECO:0000269|PubMed:18959754};
CC KM=0.29 mM for p-nitrophenyl-beta-GlcNAc (at pH 6.8 and 25 degrees
CC Celsius) {ECO:0000269|PubMed:18959754};
CC KM=0.43 mM for p-nitrophenyl-beta-GalNAc (at pH 6.8 and 25 degrees
CC Celsius) {ECO:0000269|PubMed:18959754};
CC KM=0.15 mM for p-nitrophenyl-beta-GlcNAc (at pH 6.0 and 25 degrees
CC Celsius) {ECO:0000269|PubMed:25436416};
CC KM=0.107 mM for 4-methylumbelliferone-N-acetyl-beta-D-glucosaminide
CC (4MU-beta-GlcNAc) (at 25 degrees Celsius)
CC {ECO:0000269|PubMed:21106526};
CC KM=0.148 mM for GlcNAc-beta-1,4-GlcNAc (at pH 7.0 and 25 degrees
CC Celsius) {ECO:0000269|PubMed:21106526};
CC Note=kcat is 934 sec(-1) with GlcNAc-beta-1,4-GlcNAc as substrate.
CC kcat is 708 sec(-1) with p-nitrophenyl-beta-GlcNAc as substrate. kcat
CC is 766 sec(-1) with p-nitrophenyl-beta-GalNAc as substrate
CC (PubMed:18959754). kcat is 434.7 sec(-1) with 4MU-beta-GlcNAc as
CC substrate. kcat is 507.4 sec(-1) with GlcNAc-beta-1,4-GlcNAc as
CC substrate (PubMed:21106526). {ECO:0000269|PubMed:18959754,
CC ECO:0000269|PubMed:21106526};
CC pH dependence:
CC Optimum pH is around 7.0. {ECO:0000269|PubMed:18959754};
CC Temperature dependence:
CC Optimum temperature is 30 degrees Celsius. Stable up to 40 degrees
CC Celsius. {ECO:0000269|PubMed:18959754};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:18959754,
CC ECO:0000269|PubMed:21106526, ECO:0000269|PubMed:21692744}.
CC -!- DEVELOPMENTAL STAGE: Expression is dramatically up-regulated at the
CC wandering stage. Low expression at fifth instar larva (day 1),
CC prepupal, pupal (days 1 and 2) and adult stages (PubMed:18959754).
CC Expression is up-regulated before each molting stage during development
CC of the late fourth instar larva (4L), late fifth instar larva (5L),
CC prepupa (PP), and late pupa (P3). The expression level reaches its peak
CC at the fifth instar day 5 (5L5) and prepupa stages, and the level is
CC about 10 times higher than that of fifth instar day 3 (5L3) larva. At
CC the 5L3 stage, expression levels in the integument and alimentary tract
CC are similar. However, at the 5L5 stage, the expression level in the
CC integument is up-regulated more than 3-fold, but remains unchanged in
CC the alimentary tract (PubMed:21106526). {ECO:0000269|PubMed:18959754,
CC ECO:0000269|PubMed:21106526}.
CC -!- MASS SPECTROMETRY: Mass=66719; Method=MALDI;
CC Evidence={ECO:0000269|PubMed:18959754};
CC -!- DISRUPTION PHENOTYPE: RNA interference of this gene does not cause any
CC visible changes in phenotype in larvae before pupation. At the pupation
CC stage, 20% of the larvae show abnormal phenotypes. They fail to shed
CC their old cuticles completely before starting to form new ones
CC underneath. {ECO:0000269|PubMed:21106526}.
CC -!- BIOTECHNOLOGY: A potential target for eco-friendly pesticide
CC development because of its species specificity and narrow substrate
CC spectrum (PubMed:18959754). Q2, the designed non-carbohydrate
CC naphthalimide-based inhibitor against this enzyme has potential use as
CC pharmaceutical or pesticide as it selectively recognizes chitinolytic
CC beta-GlcNAcases, but not glycoconjugate-lytic beta-GlcNAcases
CC (PubMed:25155420). {ECO:0000269|PubMed:25155420,
CC ECO:0000305|PubMed:18959754}.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 20 family.
CC {ECO:0000255|PIRNR:PIRNR001093}.
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DR EMBL; DQ887769; ABI81756.1; -; mRNA.
DR PDB; 3NSM; X-ray; 2.10 A; A=23-594.
DR PDB; 3NSN; X-ray; 2.10 A; A=23-594.
DR PDB; 3OZO; X-ray; 2.00 A; A=23-594.
DR PDB; 3OZP; X-ray; 2.00 A; A=23-594.
DR PDB; 3S6T; X-ray; 2.30 A; A=20-594.
DR PDB; 3VTR; X-ray; 2.50 A; A=23-594.
DR PDB; 3WMB; X-ray; 2.70 A; A=23-594.
DR PDB; 3WMC; X-ray; 2.10 A; A=23-594.
DR PDB; 5Y0V; X-ray; 2.42 A; A=23-594.
DR PDB; 5Y1B; X-ray; 2.21 A; A=23-594.
DR PDBsum; 3NSM; -.
DR PDBsum; 3NSN; -.
DR PDBsum; 3OZO; -.
DR PDBsum; 3OZP; -.
DR PDBsum; 3S6T; -.
DR PDBsum; 3VTR; -.
DR PDBsum; 3WMB; -.
DR PDBsum; 3WMC; -.
DR PDBsum; 5Y0V; -.
DR PDBsum; 5Y1B; -.
DR AlphaFoldDB; Q06GJ0; -.
DR SMR; Q06GJ0; -.
DR BindingDB; Q06GJ0; -.
DR ChEMBL; CHEMBL4295590; -.
DR CAZy; GH20; Glycoside Hydrolase Family 20.
DR BRENDA; 3.2.1.52; 8311.
DR EvolutionaryTrace; Q06GJ0; -.
DR GO; GO:0004563; F:beta-N-acetylhexosaminidase activity; IEA:UniProtKB-EC.
DR GO; GO:0102148; F:N-acetyl-beta-D-galactosaminidase activity; IEA:UniProtKB-EC.
DR GO; GO:0006032; P:chitin catabolic process; IEA:UniProtKB-KW.
DR GO; GO:0000272; P:polysaccharide catabolic process; IEA:UniProtKB-KW.
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; IPR029019; HEX_eukaryotic_N.
DR PANTHER; PTHR22600; PTHR22600; 1.
DR Pfam; PF00728; Glyco_hydro_20; 1.
DR Pfam; PF14845; Glycohydro_20b2; 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; Chitin degradation;
KW Direct protein sequencing; Disulfide bond; Glycoprotein; Glycosidase;
KW Hydrolase; Polysaccharide degradation; Signal.
FT SIGNAL 1..22
FT /evidence="ECO:0000255"
FT CHAIN 23..594
FT /note="Chitooligosaccharidolytic beta-N-
FT acetylglucosaminidase"
FT /evidence="ECO:0000255"
FT /id="PRO_5004165440"
FT ACT_SITE 249
FT /note="Charge relay system"
FT /evidence="ECO:0000305|PubMed:21106526"
FT ACT_SITE 303
FT /note="Charge relay system"
FT /evidence="ECO:0000305|PubMed:21106526"
FT ACT_SITE 368
FT /note="Charge relay system"
FT /evidence="ECO:0000305|PubMed:21106526"
FT SITE 327
FT /note="Important determinant of glycosidic bond
FT specificity"
FT /evidence="ECO:0000269|PubMed:23300622"
FT SITE 328
FT /note="Essential for chitooligosaccharide substrate
FT binding"
FT /evidence="ECO:0000269|PubMed:23300622"
FT SITE 490
FT /note="Essential for chitooligosaccharide substrate
FT binding"
FT /evidence="ECO:0000269|PubMed:23300622"
FT CARBOHYD 164
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498,
FT ECO:0000269|PubMed:25155420, ECO:0000269|PubMed:30135205,
FT ECO:0007744|PDB:3WMB, ECO:0007744|PDB:3WMC,
FT ECO:0007744|PDB:5Y0V, ECO:0007744|PDB:5Y1B"
FT CARBOHYD 375
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00498,
FT ECO:0000269|PubMed:25155420, ECO:0000269|PubMed:30135205,
FT ECO:0007744|PDB:3WMB, ECO:0007744|PDB:3WMC,
FT ECO:0007744|PDB:5Y0V, ECO:0007744|PDB:5Y1B"
FT DISULFID 31..59
FT /evidence="ECO:0000269|PubMed:21106526,
FT ECO:0000269|PubMed:21692744, ECO:0000269|PubMed:23300622,
FT ECO:0000269|PubMed:25155420, ECO:0000269|PubMed:25436416,
FT ECO:0000269|PubMed:30135205, ECO:0007744|PDB:3NSM,
FT ECO:0007744|PDB:3NSN, ECO:0007744|PDB:3OZO,
FT ECO:0007744|PDB:3OZP, ECO:0007744|PDB:3S6T,
FT ECO:0007744|PDB:3VTR, ECO:0007744|PDB:3WMB,
FT ECO:0007744|PDB:3WMC, ECO:0007744|PDB:5Y0V,
FT ECO:0007744|PDB:5Y1B"
FT DISULFID 36..55
FT /evidence="ECO:0000269|PubMed:21106526,
FT ECO:0000269|PubMed:21692744, ECO:0000269|PubMed:23300622,
FT ECO:0000269|PubMed:25155420, ECO:0000269|PubMed:25436416,
FT ECO:0000269|PubMed:30135205, ECO:0007744|PDB:3NSM,
FT ECO:0007744|PDB:3NSN, ECO:0007744|PDB:3OZO,
FT ECO:0007744|PDB:3OZP, ECO:0007744|PDB:3S6T,
FT ECO:0007744|PDB:3VTR, ECO:0007744|PDB:3WMB,
FT ECO:0007744|PDB:3WMC, ECO:0007744|PDB:5Y0V,
FT ECO:0007744|PDB:5Y1B"
FT DISULFID 316..373
FT /evidence="ECO:0000269|PubMed:21106526,
FT ECO:0000269|PubMed:21692744, ECO:0000269|PubMed:23300622,
FT ECO:0000269|PubMed:25155420, ECO:0000269|PubMed:25436416,
FT ECO:0000269|PubMed:30135205, ECO:0007744|PDB:3NSM,
FT ECO:0007744|PDB:3NSN, ECO:0007744|PDB:3OZO,
FT ECO:0007744|PDB:3OZP, ECO:0007744|PDB:3S6T,
FT ECO:0007744|PDB:3VTR, ECO:0007744|PDB:3WMB,
FT ECO:0007744|PDB:3WMC, ECO:0007744|PDB:5Y0V,
FT ECO:0007744|PDB:5Y1B"
FT DISULFID 326..331
FT /evidence="ECO:0000269|PubMed:21106526,
FT ECO:0000269|PubMed:21692744, ECO:0000269|PubMed:23300622,
FT ECO:0000269|PubMed:25155420, ECO:0000269|PubMed:25436416,
FT ECO:0000269|PubMed:30135205, ECO:0007744|PDB:3NSM,
FT ECO:0007744|PDB:3NSN, ECO:0007744|PDB:3OZO,
FT ECO:0007744|PDB:3OZP, ECO:0007744|PDB:3S6T,
FT ECO:0007744|PDB:3VTR, ECO:0007744|PDB:3WMB,
FT ECO:0007744|PDB:3WMC, ECO:0007744|PDB:5Y0V,
FT ECO:0007744|PDB:5Y1B"
FT DISULFID 478..491
FT /evidence="ECO:0000269|PubMed:21106526,
FT ECO:0000269|PubMed:21692744, ECO:0000269|PubMed:23300622,
FT ECO:0000269|PubMed:25155420, ECO:0000269|PubMed:25436416,
FT ECO:0000269|PubMed:30135205, ECO:0007744|PDB:3NSM,
FT ECO:0007744|PDB:3NSN, ECO:0007744|PDB:3OZO,
FT ECO:0007744|PDB:3OZP, ECO:0007744|PDB:3S6T,
FT ECO:0007744|PDB:3VTR, ECO:0007744|PDB:3WMB,
FT ECO:0007744|PDB:3WMC, ECO:0007744|PDB:5Y0V,
FT ECO:0007744|PDB:5Y1B"
FT DISULFID 585..592
FT /evidence="ECO:0000269|PubMed:21106526,
FT ECO:0000269|PubMed:21692744, ECO:0000269|PubMed:23300622,
FT ECO:0000269|PubMed:25155420, ECO:0000269|PubMed:25436416,
FT ECO:0000269|PubMed:30135205, ECO:0007744|PDB:3NSM,
FT ECO:0007744|PDB:3NSN, ECO:0007744|PDB:3OZO,
FT ECO:0007744|PDB:3OZP, ECO:0007744|PDB:3S6T,
FT ECO:0007744|PDB:3VTR, ECO:0007744|PDB:3WMB,
FT ECO:0007744|PDB:3WMC, ECO:0007744|PDB:5Y0V,
FT ECO:0007744|PDB:5Y1B"
FT MUTAGEN 327
FT /note="V->G: 5.3-fold decrease in Ki for PUGNAc inhibitor
FT as a result of widened active pocket entrance. Slight
FT decrease in KM, but decreased catalytic activity with 4MU-
FT beta-GlcNAc or GlcNAc-beta-1,4-GlcNAc as substrates. No
FT effect in sensitivity toward TMG-chitotriomycin inhibitor.
FT Able to efficiently hydrolyze GlcNAc-beta-1,2-Man in
FT contrast to the wild-type."
FT /evidence="ECO:0000269|PubMed:21106526,
FT ECO:0000269|PubMed:21692744, ECO:0000269|PubMed:23300622"
FT MUTAGEN 328
FT /note="E->A: 19% decrease in catalytic activity with 4MU-
FT beta-GlcNAc as substrate. 8-fold increase in KM for GlcNAc-
FT beta-1,4-GlcNAc. 42-fold increase in Ki for TMG-
FT chitotriomycin inhibitor. No effect in KM with p-
FT nitrophenyl-beta-GlcNAc as substrate. 9.8-fold increase in
FT Ki for NMAGT inhibitor. No effect in Ki with NGT as the
FT inhibitor."
FT /evidence="ECO:0000269|PubMed:21106526,
FT ECO:0000269|PubMed:23300622, ECO:0000269|PubMed:25436416"
FT MUTAGEN 328
FT /note="E->Q: 19% decrease in catalytic activity with 4MU-
FT beta-GlcNAc as substrate. 2.5-fold increase in KM for
FT GlcNAc-beta-1,4-GlcNAc. kcat is 12% of the wild-type value
FT using GlcNAc-beta-1,4-GlcNAc as substrate. 1.6-fold
FT increase in Ki for TMG-chitotriomycin inhibitor."
FT /evidence="ECO:0000269|PubMed:21106526,
FT ECO:0000269|PubMed:23300622"
FT MUTAGEN 433
FT /note="H->A: 1389-fold decrease in catalytic activity with
FT 4MU-beta-GlcNAc as substrate."
FT /evidence="ECO:0000269|PubMed:21106526"
FT MUTAGEN 448
FT /note="W->A: 2-fold increase in KM, 927-fold decrease in
FT kcat and a 1900-fold decrease in kcat/KM with 4MU-beta-
FT GlcNAc as substrate."
FT /evidence="ECO:0000269|PubMed:21106526"
FT MUTAGEN 448
FT /note="W->F: Slight increase in KM and more than 1000-fold
FT decrease in kcat/KM with 4MU-beta-GlcNAc as substrate."
FT /evidence="ECO:0000269|PubMed:21106526"
FT MUTAGEN 490
FT /note="W->A: 2,277-fold increase in Ki for TMG-
FT chitotriomycin inhibitor. 13-fold increase in KM for
FT GlcNAc-beta-1,4-GlcNAc. KM similar to that of wild-type,
FT but 62% decrease in catalytic activity with 4MU-beta-GlcNAc
FT as substrate. No inhibition by Q2, a synthesized non-
FT carbohydrate unsymmetrical dyad of naphthalimide and
FT thiadiazole having a dimethylamino group at C4 of the
FT naphthalimide. No inhibition by berberine."
FT /evidence="ECO:0000269|PubMed:21106526,
FT ECO:0000269|PubMed:23300622, ECO:0000269|PubMed:25155420,
FT ECO:0000269|PubMed:30135205"
FT STRAND 27..32
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 35..40
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 52..59
FT /evidence="ECO:0007829|PDB:3OZO"
FT TURN 61..64
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 79..91
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 98..112
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 113..115
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 126..137
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 151..158
FT /evidence="ECO:0007829|PDB:3OZO"
FT TURN 159..161
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 162..171
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 172..183
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 186..189
FT /evidence="ECO:0007829|PDB:3OZO"
FT TURN 190..193
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 194..205
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 210..217
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 219..221
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 225..237
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 242..246
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 258..260
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 262..267
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 268..273
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 277..289
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 293..303
FT /evidence="ECO:0007829|PDB:3OZO"
FT TURN 309..312
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 314..316
FT /evidence="ECO:0007829|PDB:3WMC"
FT HELIX 322..324
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 327..330
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 340..355
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 362..365
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 371..375
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 378..386
FT /evidence="ECO:0007829|PDB:3OZO"
FT TURN 393..395
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 396..415
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 421..427
FT /evidence="ECO:0007829|PDB:3OZO"
FT TURN 431..433
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 434..437
FT /evidence="ECO:0007829|PDB:3OZO"
FT TURN 440..442
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 443..447
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 455..461
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 465..468
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 471..474
FT /evidence="ECO:0007829|PDB:3OZO"
FT TURN 476..479
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 483..488
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 497..502
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 505..509
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 510..515
FT /evidence="ECO:0007829|PDB:3OZO"
FT STRAND 516..523
FT /evidence="ECO:0007829|PDB:3OZO"
FT TURN 530..532
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 533..537
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 540..550
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 556..558
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 560..572
FT /evidence="ECO:0007829|PDB:3OZO"
FT HELIX 583..587
FT /evidence="ECO:0007829|PDB:3OZO"
SQ SEQUENCE 594 AA; 67964 MW; 87E8DE2ADE107ED9 CRC64;
MWSRRIPLFI FGVLVLILSV AAEDVVWRWS CDNGKCVKLK NDPRSSEPAL SLEACKMFCN
EYGLLWPRPT GEADLGNFLS KINLNSIEVK ILKKGATDDL MEAAAKRFKE QVSLAIPRGS
TPKLTGKAVD VYLVNENPNE KAFSLEMDES YGLRVSPSGA DRVNATITAN SFFGMRHGLE
TLSQLFVFDD IRDHLLMVRD VNISDKPVYP YRGILLDTAR NYYSIESIKR TIEAMAAVKL
NTFHWHITDS QSFPFVTTKR PNLYKFGALS PQKVYTKAAI REVVRFGLER GVRVLPEFDA
PAHVGEGWQD TDLTVCFKAE PWKSYCVEPP CGQLNPTKDE LYQYLEDIYS DMAEVFDTTD
IFHMGGDEVS EACWNSSDSI QNFMMQNRWD LDKESFLKLW NYFQQKAQDK AYKAFGKKLP
LILWTSTLTN YKHIDDYLNK DDYIIQVWTT GVDPQIKGLL EKGYRLIMSN YDALYFDCGY
GAWVGAGNNW CSPYIGWQKV YDNSPAVIAL EHRDQVLGGE AALWSEQSDT STLDGRLWPR
AAALAERLWA EPATSWQDAE YRMLHIRERL VRMGIQAESL QPEWCYQNEG YCYS
