HGGL_SECCE
ID HGGL_SECCE Reviewed; 568 AA.
AC Q9FYS3;
DT 16-OCT-2013, integrated into UniProtKB/Swiss-Prot.
DT 01-MAR-2001, sequence version 1.
DT 03-AUG-2022, entry version 81.
DE RecName: Full=4-hydroxy-7-methoxy-3-oxo-3,4-dihydro-2H-1,4-benzoxazin-2-yl glucoside beta-D-glucosidase, chloroplastic;
DE EC=3.2.1.182;
DE AltName: Full=Beta-glucosidase;
DE Short=ScGlu;
DE EC=3.2.1.21;
DE Flags: Precursor;
OS Secale cereale (Rye).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; Liliopsida; Poales; Poaceae; BOP clade;
OC Pooideae; Triticodae; Triticeae; Hordeinae; Secale.
OX NCBI_TaxID=4550;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND TISSUE SPECIFICITY.
RC STRAIN=cv. Motto; TISSUE=Coleoptile;
RX DOI=10.1034/j.1399-3054.2003.00118.x;
RA Nikus J., Esen A., Jonsson L.M.V.;
RT "Cloning of a plastidic rye (Secale cereale) beta-glucosidase cDNA and its
RT expression in Escherichia coli.";
RL Physiol. Plantarum 118:337-345(2003).
RN [2]
RP PROTEIN SEQUENCE OF 51-70, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, ACTIVITY REGULATION, AND SUBCELLULAR LOCATION.
RC STRAIN=cv. Haru-ichiban;
RX PubMed=10773341; DOI=10.1016/s0168-9452(00)00204-1;
RA Sue M., Ishihara A., Iwamura H.;
RT "Purification and characterization of a beta-glucosidase from rye (Secale
RT cereale L.) seedlings.";
RL Plant Sci. 155:67-74(2000).
RN [3]
RP SUBUNIT, BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS OF GLU-240;
RP PHE-247; TYR-427; GLU-456; GLY-513; SER-514 AND PHE-520.
RX PubMed=16751439; DOI=10.1104/pp.106.077693;
RA Sue M., Yamazaki K., Yajima S., Nomura T., Matsukawa T., Iwamura H.,
RA Miyamoto T.;
RT "Molecular and structural characterization of hexameric beta-D-glucosidases
RT in wheat and rye.";
RL Plant Physiol. 141:1237-1247(2006).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) OF 50-568 IN COMPLEX WITH DIMBOA OR
RP INHIBITOR.
RX PubMed=21421370; DOI=10.1016/j.plantsci.2010.09.001;
RA Sue M., Nakamura C., Miyamoto T., Yajima S.;
RT "Active-site architecture of benzoxazinone-glucoside beta-D-glucosidases in
RT Triticeae.";
RL Plant Sci. 180:268-275(2011).
CC -!- FUNCTION: Involved in defense of young plant parts against pests via
CC the production of benzoxazolinones (hydroxamic acids) from hydroxamic
CC acid glucosides. The preferred substrate is DIBOA-beta-D-glucoside. Can
CC also use esculin and genistein glucoside as substrates, but no activity
CC with salicin, p-nitrophenyl-alpha-glucoside or substrates related to
CC cell wall components. {ECO:0000269|PubMed:10773341, ECO:0000269|Ref.1}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=DIMBOA beta-D-glucoside + H2O = D-glucose + DIMBOA;
CC Xref=Rhea:RHEA:33975, ChEBI:CHEBI:4167, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:18048, ChEBI:CHEBI:37573; EC=3.2.1.182;
CC -!- CATALYTIC ACTIVITY:
CC Reaction=DIBOA beta-D-glucoside + H2O = D-glucose + DIBOA;
CC Xref=Rhea:RHEA:33979, ChEBI:CHEBI:4167, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:63558, ChEBI:CHEBI:63670; EC=3.2.1.182;
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Hydrolysis of terminal, non-reducing beta-D-glucosyl residues
CC with release of beta-D-glucose.; EC=3.2.1.21;
CC -!- ACTIVITY REGULATION: Inhibited by castanospermine, Ag(+) and Cu(2+).
CC 34% inhibition by Zn(2+) and not affected by EDTA.
CC {ECO:0000269|PubMed:10773341}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.19 mM for DIBOA-beta-D-glucoside {ECO:0000269|PubMed:10773341};
CC KM=3.3 mM for DIBOA-beta-D-glucoside {ECO:0000269|Ref.1};
CC KM=0.8 mM for DIBOA-beta-D-glucoside (with recombinant enzyme)
CC {ECO:0000269|PubMed:16751439};
CC KM=0.617 mM for DIMBOA-beta-D-glucoside
CC {ECO:0000269|PubMed:10773341};
CC KM=2.1 mM for DIMBOA-beta-D-glucoside {ECO:0000269|Ref.1};
CC KM=1.3 mM for DIMBOA-beta-D-glucoside (with recombinant enzyme)
CC {ECO:0000269|PubMed:16751439};
CC KM=2.0 mM for HBOA-beta-D-glucoside {ECO:0000269|PubMed:10773341};
CC KM=0.893 mM for HMBOA-beta-D-glucoside {ECO:0000269|PubMed:10773341};
CC KM=1.4 mM for o-nitrophenyl beta-D-glucopyranoside
CC {ECO:0000269|Ref.1};
CC KM=0.9 mM for p-nitrophenyl beta-D-glucopyranoside
CC {ECO:0000269|PubMed:10773341};
CC KM=2.4 mM for p-nitrophenyl beta-D-glucopyranoside
CC {ECO:0000269|Ref.1};
CC KM=1.78 mM for p-nitrophenyl beta-D-glucopyranoside (with recombinant
CC enzyme) {ECO:0000269|PubMed:16751439};
CC KM=1.3 mM for p-nitrophenyl beta-D-galactopyranoside
CC {ECO:0000269|Ref.1};
CC KM=3.17 mM for p-nitrophenyl beta-D-xyloside
CC {ECO:0000269|PubMed:10773341};
CC KM=0.616 mM for p-nitrophenyl beta-D-fucoside
CC {ECO:0000269|PubMed:10773341};
CC KM=0.8 mM for p-nitrophenyl beta-D-fucoside {ECO:0000269|Ref.1};
CC KM=0.151 mM for esculin {ECO:0000269|PubMed:10773341};
CC Vmax=5870 nmol/sec/mg enzyme with DIBOA-beta-D-glucoside as substrate
CC {ECO:0000269|PubMed:10773341};
CC Vmax=2567 nmol/sec/mg enzyme with DIBOA-beta-D-glucoside as substrate
CC {ECO:0000269|Ref.1};
CC Vmax=4952 nmol/sec/mg enzyme with DIMBOA-beta-D-glucoside as
CC substrate {ECO:0000269|PubMed:10773341};
CC Vmax=1786 nmol/sec/mg enzyme with DIMBOA-beta-D-glucoside as
CC substrate {ECO:0000269|Ref.1};
CC Vmax=2375 nmol/sec/mg enzyme with o-nitrophenyl beta-D-
CC glucopyranoside as substrate {ECO:0000269|Ref.1};
CC Vmax=1420 nmol/sec/mg enzyme with HBOA-beta-D-glucoside as substrate
CC {ECO:0000269|PubMed:10773341};
CC Vmax=1005 nmol/sec/mg enzyme with HMBOA-beta-D-glucoside as substrate
CC {ECO:0000269|PubMed:10773341};
CC Vmax=828 nmol/sec/mg enzyme with p-nitrophenyl beta-D-glucopyranoside
CC as substrate {ECO:0000269|PubMed:10773341};
CC Vmax=779 nmol/sec/mg enzyme with p-nitrophenyl beta-D-glucopyranoside
CC as substrate {ECO:0000269|Ref.1};
CC Vmax=78.2 nmol/sec/mg enzyme with p-nitrophenyl beta-D-xyloside as
CC substrate {ECO:0000269|PubMed:10773341};
CC Vmax=1671 nmol/sec/mg enzyme with p-nitrophenyl beta-D-fucoside as
CC substrate {ECO:0000269|PubMed:10773341};
CC Vmax=997 nmol/sec/mg enzyme with p-nitrophenyl beta-D-fucoside as
CC substrate {ECO:0000269|Ref.1};
CC Vmax=51 nmol/sec/mg enzyme with p-nitrophenyl beta-D-
CC galactopyranoside as substrate {ECO:0000269|Ref.1};
CC Vmax=974 nmol/sec/mg enzyme with esculin as substrate
CC {ECO:0000269|PubMed:10773341};
CC Note=kcat is 172 sec(-1) with DIBOA-beta-D-glucoside as substrate.
CC kcat is 118 sec(-1) with DIBOA-beta-D-glucoside as substrate (with
CC recombinant enzyme). kcat is 119 sec(-1) with DIMBOA-beta-D-glucoside
CC as substrate. kcat is 158 sec(-1) with DIMBOA-beta-D-glucoside as
CC substrate (with recombinant enzyme). kcat is 159 sec(-1) with o-
CC nitrophenyl beta-D-glucopyranoside as substrate. kcat is 66 sec(-1)
CC with p-nitrophenyl beta-D-fucoside as substrate. kcat is 52 sec(-1)
CC with p-nitrophenyl beta-D-glucopyranoside as substrate. kcat is 3.5
CC sec(-1) with p-nitrophenyl beta-D-galactopyranoside as substrate.
CC kcat is 22.2 sec(-1) with p-nitrophenyl beta-D-glucopyranoside as
CC substrate (with recombinant enzyme).;
CC pH dependence:
CC Optimum pH is 5.5. {ECO:0000269|PubMed:10773341,
CC ECO:0000269|PubMed:16751439, ECO:0000269|Ref.1};
CC Temperature dependence:
CC Optimum temperature is 25-30 degrees Celsius.
CC {ECO:0000269|PubMed:10773341, ECO:0000269|PubMed:16751439,
CC ECO:0000269|Ref.1};
CC -!- SUBUNIT: Homohexamer. {ECO:0000269|PubMed:16751439,
CC ECO:0000269|PubMed:21421370}.
CC -!- SUBCELLULAR LOCATION: Plastid, chloroplast {ECO:0000305}.
CC -!- TISSUE SPECIFICITY: Expressed in seedlings, mesocotyl, coleoptile, leaf
CC sheath, and roots. {ECO:0000269|Ref.1}.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 1 family. {ECO:0000305}.
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DR EMBL; AF293849; AAG00614.1; -; mRNA.
DR PDB; 3AIU; X-ray; 2.20 A; A=50-568.
DR PDB; 3AIV; X-ray; 2.50 A; A=50-568.
DR PDB; 3AIW; X-ray; 2.40 A; A=50-568.
DR PDBsum; 3AIU; -.
DR PDBsum; 3AIV; -.
DR PDBsum; 3AIW; -.
DR AlphaFoldDB; Q9FYS3; -.
DR SMR; Q9FYS3; -.
DR CAZy; GH1; Glycoside Hydrolase Family 1.
DR BRENDA; 3.2.1.182; 5654.
DR BRENDA; 3.2.1.21; 5654.
DR SABIO-RK; Q9FYS3; -.
DR EvolutionaryTrace; Q9FYS3; -.
DR GO; GO:0009507; C:chloroplast; IEA:UniProtKB-SubCell.
DR GO; GO:0008422; F:beta-glucosidase activity; IEA:UniProtKB-EC.
DR GO; GO:0102726; F:DIMBOA glucoside beta-D-glucosidase activity; IEA:UniProtKB-EC.
DR GO; GO:0102483; F:scopolin beta-glucosidase activity; IEA:UniProtKB-EC.
DR GO; GO:0005975; P:carbohydrate metabolic process; IEA:InterPro.
DR InterPro; IPR001360; Glyco_hydro_1.
DR InterPro; IPR018120; Glyco_hydro_1_AS.
DR InterPro; IPR033132; Glyco_hydro_1_N_CS.
DR InterPro; IPR017853; Glycoside_hydrolase_SF.
DR PANTHER; PTHR10353; PTHR10353; 1.
DR Pfam; PF00232; Glyco_hydro_1; 1.
DR PRINTS; PR00131; GLHYDRLASE1.
DR SUPFAM; SSF51445; SSF51445; 1.
DR PROSITE; PS00572; GLYCOSYL_HYDROL_F1_1; 1.
DR PROSITE; PS00653; GLYCOSYL_HYDROL_F1_2; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Chloroplast; Direct protein sequencing; Disulfide bond;
KW Glycosidase; Hydrolase; Plastid; Transit peptide.
FT TRANSIT 1..50
FT /note="Chloroplast"
FT /evidence="ECO:0000269|PubMed:10773341"
FT CHAIN 51..568
FT /note="4-hydroxy-7-methoxy-3-oxo-3,4-dihydro-2H-1,4-
FT benzoxazin-2-yl glucoside beta-D-glucosidase,
FT chloroplastic"
FT /id="PRO_0000424096"
FT ACT_SITE 240
FT /note="Proton donor"
FT /evidence="ECO:0000250"
FT ACT_SITE 456
FT /note="Nucleophile"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10055"
FT BINDING 92
FT /ligand="substrate"
FT BINDING 194
FT /ligand="substrate"
FT BINDING 239
FT /ligand="substrate"
FT BINDING 511..512
FT /ligand="substrate"
FT DISULFID 259..265
FT /evidence="ECO:0000250"
FT MUTAGEN 240
FT /note="E->A: Total loss of activity."
FT /evidence="ECO:0000269|PubMed:16751439"
FT MUTAGEN 247
FT /note="F->A: 96% loss of activity with DIBOA-Glc and
FT DIMBOA-Glc."
FT /evidence="ECO:0000269|PubMed:16751439"
FT MUTAGEN 427
FT /note="Y->A: 7% loss of activity with DIBOA-Glc and 4.8
FT fold increased activity with DIMBOA-Glc."
FT /evidence="ECO:0000269|PubMed:16751439"
FT MUTAGEN 427
FT /note="Y->F: 87% loss of activity with DIBOA-Glc and 1.5
FT fold increased activity with DIMBOA-Glc."
FT /evidence="ECO:0000269|PubMed:16751439"
FT MUTAGEN 456
FT /note="E->A: Total loss of activity."
FT /evidence="ECO:0000269|PubMed:16751439"
FT MUTAGEN 513
FT /note="G->F: 77% loss of activity with DIBOA-Glc and 81%
FT with DIMBOA-Glc."
FT /evidence="ECO:0000269|PubMed:16751439"
FT MUTAGEN 513
FT /note="G->S: 42% loss of activity with DIBOA-Glc and 2 fold
FT increased activity with DIMBOA-Glc. 42% loss of activity
FT with DIBOA-Glc and 4 fold increased activity with DIMBOA-
FT Glc; when associated with L-514."
FT /evidence="ECO:0000269|PubMed:16751439"
FT MUTAGEN 514
FT /note="S->L: 62% loss of activity with DIBOA-Glc and 2.4
FT fold increased activity with DIMBOA-Glc. 42% loss of
FT activity with DIBOA-Glc and 4 fold increased activity with
FT DIMBOA-Glc; when associated with S-513."
FT /evidence="ECO:0000269|PubMed:16751439"
FT MUTAGEN 520
FT /note="F->Y: 19% loss of activity with DIBOA-Glc and 2.5
FT fold increased activity with DIMBOA-Glc."
FT /evidence="ECO:0000269|PubMed:16751439"
FT HELIX 69..71
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 75..77
FT /evidence="ECO:0007829|PDB:3AIU"
FT STRAND 83..87
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 90..93
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 106..113
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 115..117
FT /evidence="ECO:0007829|PDB:3AIU"
FT STRAND 124..126
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 130..144
FT /evidence="ECO:0007829|PDB:3AIU"
FT STRAND 147..152
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 155..158
FT /evidence="ECO:0007829|PDB:3AIU"
FT STRAND 162..165
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 168..183
FT /evidence="ECO:0007829|PDB:3AIU"
FT STRAND 187..195
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 199..205
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 207..209
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 212..228
FT /evidence="ECO:0007829|PDB:3AIU"
FT TURN 229..231
FT /evidence="ECO:0007829|PDB:3AIU"
FT STRAND 234..239
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 241..249
FT /evidence="ECO:0007829|PDB:3AIU"
FT STRAND 265..267
FT /evidence="ECO:0007829|PDB:3AIU"
FT TURN 272..274
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 275..297
FT /evidence="ECO:0007829|PDB:3AIU"
FT STRAND 305..321
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 322..335
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 337..345
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 350..356
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 357..359
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 365..371
FT /evidence="ECO:0007829|PDB:3AIU"
FT STRAND 376..390
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 402..406
FT /evidence="ECO:0007829|PDB:3AIU"
FT STRAND 408..413
FT /evidence="ECO:0007829|PDB:3AIU"
FT STRAND 419..421
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 433..445
FT /evidence="ECO:0007829|PDB:3AIU"
FT STRAND 452..456
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 475..493
FT /evidence="ECO:0007829|PDB:3AIU"
FT STRAND 498..504
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 512..517
FT /evidence="ECO:0007829|PDB:3AIU"
FT STRAND 522..525
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 527..529
FT /evidence="ECO:0007829|PDB:3AIU"
FT STRAND 533..535
FT /evidence="ECO:0007829|PDB:3AIU"
FT HELIX 537..545
FT /evidence="ECO:0007829|PDB:3AIU"
SQ SEQUENCE 568 AA; 64212 MW; 11F7E87C0CB81DF7 CRC64;
MALLVGGTLN PTTHLSLRSR AGRNSENVWL RSAASSQTSK GRFCNLTVRA GTPSKPSEPI
GPVFTKLKPW QIPKRDWFSK DFLFGASTSA YQIEGAWNED GKGPSTWDHF CHTYPERISD
GTNGDVAANS YHMYEEDVKA LKDMGMKVYR FSISWSRILP NGTGKPNQKG IDYYNNLINS
LIRHGIVPYV TIWHWDTPQA LEDKYGGFLD KQIVNDYKYF AELCFQSFGD RVKNWFTFNE
PHTYCCFSYG EGIHAPGRCS PGLDCAVPEG DSLREPYTAG HHILLAHAEA VELFKAHYNK
HGDSKIGMAF DVMGYEPYQD SFLDDQARER SIDYNMGWFL EPVVRGDYPF SMRSLIGDRL
PMFTKEEQEK LGSLCDIMGL NYYTSRFSKH VDISSDYTPT LNTDDAYASS ETTGSDGNEI
GPITGTYWIY MYPKGLTDLL LIMKEKYGNP PIFITENGIA DVEGDPEMPD PLDDWKRLDY
LQRHISAVKD AIDQGADVRG HFTWGLIDNF EWGSGYSSRF GLVYIDKEDG NKRKLKKSAK
WFAKFNSVPK TLLKTTNNNA TVTASVSV
