GFT_ALLCE
ID GFT_ALLCE Reviewed; 612 AA.
AC P92916;
DT 13-NOV-2007, integrated into UniProtKB/Swiss-Prot.
DT 01-MAY-1997, sequence version 1.
DT 03-AUG-2022, entry version 88.
DE RecName: Full=Bifunctional 6(G)-fructosyltransferase/2,1-fructan:2,1-fructan 1-fructosyltransferase;
DE Short=1-FFT;
DE Short=6G-FFT;
DE Short=6GFT;
DE Short=FFT;
DE EC=2.4.1.100 {ECO:0000269|PubMed:15720662};
DE EC=2.4.1.243 {ECO:0000269|PubMed:15720662};
OS Allium cepa (Onion).
OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC Spermatophyta; Magnoliopsida; Liliopsida; Asparagales; Amaryllidaceae;
OC Allioideae; Allieae; Allium.
OX NCBI_TaxID=4679;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], SUBCELLULAR LOCATION, AND INDUCTION.
RX PubMed=9107030; DOI=10.1046/j.1365-313x.1997.11030387.x;
RA Vijn I., van Dijken A., Sprenger N., van Dun K., Weisbeek P., Wiemken A.,
RA Smeekens S.;
RT "Fructan of the inulin neoseries is synthesized in transgenic chicory
RT plants (Cichorium intybus L.) harbouring onion (Allium cepa L.)
RT fructan:fructan 6G-fructosyltransferase.";
RL Plant J. 11:387-398(1997).
RN [2]
RP PROTEIN SEQUENCE OF 56-110 AND 460-518, FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND INHIBITION.
RC STRAIN=cv. Tenshin;
RX PubMed=15720662; DOI=10.1111/j.1469-8137.2004.01231.x;
RA Fujishima M., Sakai H., Ueno K., Takahashi N., Onodera S., Benkeblia N.,
RA Shiomi N.;
RT "Purification and characterization of a fructosyltransferase from onion
RT bulbs and its key role in the synthesis of fructo-oligosaccharides in
RT vivo.";
RL New Phytol. 165:513-524(2005).
CC -!- FUNCTION: Involved in the synthesis of fructan of the inulin neoseries.
CC Catalyzes a self-transfer between identical oligosaccharides of the 1-
CC kestose series. {ECO:0000269|PubMed:15720662}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=[1-beta-D-fructofuranosyl-(2->1)-]m+1 alpha-D-glucopyranoside
CC + [1-beta-D-fructofuranosyl-(2->1)-]n+1 alpha-D-glucopyranoside = [1-
CC beta-D-fructofuranosyl-(2->1)-]m alpha-D-glucopyranoside + [1-beta-D-
CC fructofuranosyl-(2->1)-]n+1 beta-D-fructofuranosyl-(2->6)-alpha-D-
CC glucopyranoside (m > 0, n >= 0).; EC=2.4.1.243;
CC Evidence={ECO:0000269|PubMed:15720662};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=[beta-D-fructosyl-(2->1)-]m + [beta-D-fructosyl-(2->1)-]n =
CC [beta-D-fructosyl-(2->1)-]m-1 + [beta-D-fructosyl-(2->1)-]n+1.;
CC EC=2.4.1.100; Evidence={ECO:0000269|PubMed:15720662};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=88 mM for 1-kestose (3a) (for 6G-FFT activity)
CC {ECO:0000269|PubMed:15720662};
CC KM=18 mM for 1-kestose (3a) (for 1-FFT activity)
CC {ECO:0000269|PubMed:15720662};
CC KM=310 mM for nystose (4a) (for 6G-FFT activity)
CC {ECO:0000269|PubMed:15720662};
CC KM=440 mM for nystose (4a) (for 1-FFT activity)
CC {ECO:0000269|PubMed:15720662};
CC Vmax=10.5 mmol/min/mg enzyme toward 1-kestose (3a) for 6G-FFT
CC activity {ECO:0000269|PubMed:15720662};
CC Vmax=7.51 mmol/min/mg enzyme toward 1-kestose (3a) for 1-FFT activity
CC {ECO:0000269|PubMed:15720662};
CC Vmax=34.9 mmol/min/mg enzyme toward nystose (4a) for 6G-FFT activity
CC {ECO:0000269|PubMed:15720662};
CC Vmax=3.66 mmol/min/mg enzyme toward nystose (4a) for 1-FFT activity
CC {ECO:0000269|PubMed:15720662};
CC pH dependence:
CC Optimum pH is 5.68 for both catalytic activities. Stable between pH
CC 5.0-6.3. {ECO:0000269|PubMed:15720662};
CC Temperature dependence:
CC Stable up to 40 degrees Celsius for both catalytic activities.
CC {ECO:0000269|PubMed:15720662};
CC -!- SUBCELLULAR LOCATION: Vacuole membrane {ECO:0000305|PubMed:9107030};
CC Single-pass type II membrane protein {ECO:0000305|PubMed:9107030}.
CC -!- INDUCTION: 6G-FFT activity is induced by high sucrose under continuous
CC light. Both 6G-fFFT and 1-FFT activities are strongly inhibited by
CC HgCl(2), AgNO(3), p-chloromercuribenzoate and SDS, partially inhibited
CC by CaCl(2), MgCl(2), MnCl(2), FeCl(2), CoCl(2), ZnCl(2), SnCl(2),
CC CuSO(4) and EDTA, and activated by sodium deoxycholate, triton X-100
CC and tween-80. {ECO:0000269|PubMed:9107030}.
CC -!- PTM: Might be processed in two N-terminal and C-terminal proteolytic
CC fragments.
CC -!- SIMILARITY: Belongs to the glycosyl hydrolase 32 family. {ECO:0000305}.
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DR EMBL; Y07838; CAA69170.1; -; mRNA.
DR AlphaFoldDB; P92916; -.
DR SMR; P92916; -.
DR CAZy; GH32; Glycoside Hydrolase Family 32.
DR BRENDA; 2.4.1.243; 248.
DR GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW.
DR GO; GO:0005774; C:vacuolar membrane; IEA:UniProtKB-SubCell.
DR GO; GO:0047207; F:1,2-beta-fructan 1F-fructosyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0033841; F:6G-fructosyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0004553; F:hydrolase activity, hydrolyzing O-glycosyl compounds; IEA:InterPro.
DR GO; GO:0005975; P:carbohydrate metabolic process; IEA:InterPro.
DR Gene3D; 2.115.10.20; -; 1.
DR InterPro; IPR013320; ConA-like_dom_sf.
DR InterPro; IPR001362; Glyco_hydro_32.
DR InterPro; IPR013189; Glyco_hydro_32_C.
DR InterPro; IPR013148; Glyco_hydro_32_N.
DR InterPro; IPR023296; Glyco_hydro_beta-prop_sf.
DR Pfam; PF08244; Glyco_hydro_32C; 1.
DR Pfam; PF00251; Glyco_hydro_32N; 1.
DR SMART; SM00640; Glyco_32; 1.
DR SUPFAM; SSF49899; SSF49899; 1.
DR SUPFAM; SSF75005; SSF75005; 1.
PE 1: Evidence at protein level;
KW Direct protein sequencing; Disulfide bond; Glycoprotein; Glycosidase;
KW Hydrolase; Membrane; Signal-anchor; Transferase; Transmembrane;
KW Transmembrane helix; Vacuole.
FT CHAIN 1..612
FT /note="Bifunctional 6(G)-fructosyltransferase/2,1-
FT fructan:2,1-fructan 1-fructosyltransferase"
FT /id="PRO_0000310733"
FT TOPO_DOM 1..24
FT /note="Cytoplasmic"
FT /evidence="ECO:0000255"
FT TRANSMEM 25..45
FT /note="Helical; Signal-anchor for type II membrane protein"
FT /evidence="ECO:0000255"
FT TOPO_DOM 46..612
FT /note="Vacuolar"
FT /evidence="ECO:0000255"
FT ACT_SITE 85
FT /evidence="ECO:0000250"
FT BINDING 82..85
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 101
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 109
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 144..145
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 208..209
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 267
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT CARBOHYD 111
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT CARBOHYD 216
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT CARBOHYD 230
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT CARBOHYD 465
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT CARBOHYD 586
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT CARBOHYD 603
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT DISULFID 466..514
FT /evidence="ECO:0000250"
FT CONFLICT 56
FT /note="V -> A (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 468
FT /note="S -> T (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 474
FT /note="I -> T (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 505
FT /note="I -> T (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 508
FT /note="S -> A (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 510
FT /note="I -> R (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 612 AA; 68631 MW; 6BE93637A9EAA7EA CRC64;
MDAQDIESRH PLIGARPRRR ALRSLSILLA AALLLGLVLF YANGTGSGTA VDPVRVDNEF
PWTNDMLAWQ RCGFHFRTVR NYMNDPSGPM YYKGWYHLFY QHNKDFAYWG NITWGHAVSR
DLINWQHLPV AVGPDHWYDI SGVWTGSIIV VSEDRVVMLF TGGTKSFDQS INLAEAADPS
DPLLLKWIKY DNNPILWPPP GIVRDDFRDP NPIWYNASES TYHIVVGSKN DSLQHTGIAL
VYLTKDFKKF DLLPTVLHSV DKVGMWECVE VYPVATTGPL LHKAIDNFDV DRVLDRSTVK
HVLKASMNDE WHDYYAIGTF DPIGNKWTPD DETVDVGIGL RYDWGKFYAS RTFFDPLKQR
RIIWGYIGEV DSQKADIAKG WASLQGIPRS VLYDVKTGTN VLTWPIEEME GLRMARKDFS
GIKIKKGSTV ELSDFGDAFQ IDIEAEFTIS KEALEATIEA DVGYNCSSSG GAAIRGTLGP
FGLLVLANQD LTENTATYFY VSKGIDGSLI THFCQDETRS SKANDIVKRV VGGTVPVLDG
ETFAVRILVD HSVIESFAMG GRTSATSRAY PTEAINSAAR VFLFNNATGV DVIAESVKIW
QMNSTYNDFY HF
