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Hydrolase PDB-id
1st8
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Protein chain
537 a.a. *
Ligands
NDG-NAG-MAN
NAG-NAG
GOL ×4
Waters ×331

* Residue conservation analysis
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PDB id: 1st8
Name: Hydrolase
Title: Crystal structure of fructan 1-exohydrolase iia from cichorium intybus

Structure:
Fructan 1-exohydrolase iia. Chain: a. Engineered: yes

Source:
Cichorium intybus. Chicory. Organism_taxid: 13427. Gene: 1-feh iia. Expressed in: pichia pastoris. Expression_system_taxid: 4922

UniProt:
Q93X60 (Q93X60_CICIN) Pfam   ArchSchema ?
Seq:
Struc:
Seq:
Struc:
Seq: 581 a.a.
Struc: 537 a.a.
Key:    PfamA domain
 Secondary structure  CATH domain

Enzyme class:
E.C.3.2.1.80   [IntEnz]   [ExPASy]   [KEGG]   [BRENDA]

Reaction:
Hydrolysis of terminal, non-reducing 2,1- and 2,6-linked beta-D-fructofuranose residues in fructans.

Pathway:
Fructosidase

Resolution:
2.35Å

R-factor:
0.183

R-free:
0.200

Authors:
M.Verhaest,W.Van Den Ende,C.J.De Ranter,A.Van Laere, A.Rabijns

Key ref:
M.Verhaest et al. (2005). X-ray diffraction structure of a plant glycosyl hydrolase family 32 protein: fructan 1-exohydrolase IIa of Cichorium intybus.. Plant J, 41, 400-411. [PubMed id: 15659099] [DOI: 10.1111/j.1365-313X.2004.02304.x]

Date:
25-Mar-04

Release date:
01-Mar-05
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    Key reference    
 
 
DOI no: 10.1111/j.1365-313X.2004.02304.x Plant J 41:400-411 (2005)
PubMed id: 15659099  
 
 
X-ray diffraction structure of a plant glycosyl hydrolase family 32 protein: fructan 1-exohydrolase IIa of Cichorium intybus.
M.Verhaest, W.V.Ende, K.L.Roy, C.J.De Ranter, A.V.Laere, A.Rabijns.
 
  ABSTRACT  
 
Fructan 1-exohydrolase, an enzyme involved in fructan degradation, belongs to the glycosyl hydrolase family 32. The structure of isoenzyme 1-FEH IIa from Cichorium intybus is described at a resolution of 2.35 A. The structure consists of an N-terminal fivefold beta-propeller domain connected to two C-terminal beta-sheets. The putative active site is located entirely in the beta-propeller domain and is formed by amino acids which are highly conserved within glycosyl hydrolase family 32. The fructan-binding site is thought to be in the cleft formed between the two domains. The 1-FEH IIa structure is compared with the structures of two homologous but functionally different enzymes: a levansucrase from Bacillus subtilis (glycosyl hydrolase family 68) and an invertase from Thermotoga maritima (glycosyl hydrolase family 32).
 

Literature references that cite this PDB file's key reference

  PubMed id Reference
19936386 A.Homann, and J.Seibel (2009).
Chemo-enzymatic synthesis and functional analysis of natural and modified glycostructures.
  Nat Prod Rep, 26, 1555-1571.  
18821058 D.Altenbach, E.Rudiño-Pinera, C.Olvera, T.Boller, A.Wiemken, and T.Ritsema (2009).
An acceptor-substrate binding site determining glycosyl transfer emerges from mutant analysis of a plant vacuolar invertase and a fructosyltransferase.
  Plant Mol Biol, 69, 47-56.  
19269996 K.Tamura, A.Kawakami, Y.Sanada, K.Tase, T.Komatsu, and M.Yoshida (2009).
Cloning and functional analysis of a fructosyltransferase cDNA for synthesis of highly polymerized levans in timothy (Phleum pratense L.).
  J Exp Bot, 60, 893-905.  
19726634 L.Schroeven, W.Lammens, A.Kawakami, M.Yoshida, A.Van Laere, and W.Van den Ende (2009).
Creating S-type characteristics in the F-type enzyme fructan:fructan 1-fructosyltransferase of Triticum aestivum L.
  J Exp Bot, 60, 3687-3696.  
19129163 W.Lammens, K.Le Roy, L.Schroeven, A.Van Laere, A.Rabijns, and W.Van den Ende (2009).
Structural insights into glycoside hydrolase family 32 and 68 enzymes: functional implications.
  J Exp Bot, 60, 727-740.  
19765078 W.Van den Ende, W.Lammens, A.Van Laere, L.Schroeven, and K.Le Roy (2009).
Donor and acceptor substrate selectivity among plant glycoside hydrolase family 32 enzymes.
  FEBS J, 276, 5788-5798.  
18366639 G.Meng, and K.Fütterer (2008).
Donor substrate recognition in the raffinose-bound E342A mutant of fructosyltransferase Bacillus subtilis levansucrase.
  BMC Struct Biol, 8, 16.
PDB codes: 3byj 3byk 3byl 3byn
18346102 H.Hisano, A.Kanazawa, M.Yoshida, M.O.Humphreys, M.Iizuka, K.Kitamura, and T.Yamada (2008).
Coordinated expression of functionally diverse fructosyltransferase genes is associated with fructan accumulation in response to low temperature in perennial ryegrass.
  New Phytol, 178, 766-780.  
17963237 J.Mátrai, W.Lammens, A.Jonckheer, K.Le Roy, A.Rabijns, W.Van den Ende, and M.De Maeyer (2008).
An alternate sucrose binding mode in the E203Q Arabidopsis invertase mutant: an X-ray crystallography and docking study.
  Proteins, 71, 552-564.
PDB code: 2oxb
18331426 K.Le Roy, W.Lammens, A.Van Laere, and W.Van den Ende (2008).
Influencing the binding configuration of sucrose in the active sites of chicory fructan 1-exohydrolase and sugar beet fructan 6-exohydrolase.
  New Phytol, 178, 572-580.  
17293485 C.Goosen, X.L.Yuan, J.M.van Munster, A.F.Ram, M.J.van der Maarel, and L.Dijkhuizen (2007).
Molecular and biochemical characterization of a novel intracellular invertase from Aspergillus niger with transfructosylating activity.
  Eukaryot Cell, 6, 674-681.  
17888113 K.Le Roy, M.Verhaest, A.Rabijns, S.Clerens, A.Van Laere, and W.Van den Ende (2007).
N-glycosylation affects substrate specificity of chicory fructan 1-exohydrolase: evidence for the presence of an inulin binding cleft.
  New Phytol, 176, 317-324.  
17335500 M.Verhaest, W.Lammens, K.Le Roy, C.J.De Ranter, A.Van Laere, A.Rabijns, and W.Van den Ende (2007).
Insights into the fine architecture of the active site of chicory fructan 1-exohydrolase: 1-kestose as substrate vs sucrose as inhibitor.
  New Phytol, 174, 90.
PDB codes: 2add 2ade 2aey 2aez
17192265 W.S.Jung, C.K.Hong, S.Lee, C.S.Kim, S.J.Kim, S.I.Kim, and S.Rhee (2007).
Structural and functional insights into intramolecular fructosyl transfer by inulin fructotransferase.
  J Biol Chem, 282, 8414-8423.
PDB codes: 2inu 2inv
16899050 L.K.Ozimek, S.Kralj, T.Kaper, M.J.van der Maarel, and L.Dijkhuizen (2006).
Single amino acid residue changes in subsite -1 of inulosucrase from Lactobacillus reuteri 121 strongly influence the size of products synthesized.
  FEBS J, 273, 4104-4113.  
17139091 M.Verhaest, W.Lammens, K.Le Roy, B.De Coninck, C.J.De Ranter, A.Van Laere, W.Van den Ende, and A.Rabijns (2006).
X-ray diffraction structure of a cell-wall invertase from Arabidopsis thaliana.
  Acta Crystallogr D Biol Crystallogr, 62, 1555-1563.
PDB code: 2ac1
17018033 T.Ritsema, L.Hernández, A.Verhaar, D.Altenbach, T.Boller, A.Wiemken, and S.Smeekens (2006).
Developing fructan-synthesizing capability in a plant invertase via mutations in the sucrose-binding box.
  Plant J, 48, 228-237.  
17173633 J.Chalmers, A.Lidgett, N.Cummings, Y.Cao, J.Forster, and G.Spangenberg (2005).
Molecular genetics of fructan metabolism in perennial ryegrass.
  Plant Biotechnol J, 3, 459-474.  
16158237 T.Ritsema, A.Verhaar, I.Vijn, and S.Smeekens (2005).
Using natural variation to investigate the function of individual amino acids in the sucrose-binding box of fructan:fructan 6G-fructosyltransferase (6G-FFT) in product formation.
  Plant Mol Biol, 58, 597-607.  
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time. Where a reference describes a PDB structure, the PDB codes are shown on the right.