PDBsum entry: 1eo5

Transferase

PDB-id

1eo5

Contents

Description

Header details

Protein chain

Ligands

BGC-GLC-GLC-GLC-
GLC-GLC-GLC

GLC-GLC-GLC-GLC ×2

GLC-GLC-GLC

MPD

Metal ions

_CA ×3

Waters ×674

* Residue conservation analysis

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PDB id:

1eo5

Name:

Transferase

Title:

Bacillus circulans strain 251 cyclodextrin glycosyltransferase in complex with maltoheptaose

Structure:

Protein (cyclodextrin glycosyltransferase). Chain: a. Engineered: yes. Mutation: yes

Source:

Bacillus circulans. Organism_taxid: 1397. Strain: 251. Cellular_location: extracellular. Expressed in: bacillus subtilis. Expression_system_taxid: 1423.

UniProt:

P43379 (CDGT2_BACCI)

Seq:

Struc:

Seq:

Struc:

Seq:

713 a.a.

Struc:

686 a.a.*

Key:		PfamA domain
		Secondary structure			CATH domain

* PDB and UniProt seqs differ at 3 residue positions (black crosses)

Enzyme class:

E.C.2.4.1.19 [IntEnz] [ExPASy] [KEGG] [BRENDA]

Reaction:

Degrades starch to cyclodextrins by formation of a 1,4-alpha-D- glucosidic bond.

Resolution:

2.00Å

R-factor:

0.169

R-free:

0.210

Authors:

J.C.M.Uitdehaag,B.W.Dijkstra

Key ref:

J.C.Uitdehaag et al. (2000). Structures of maltohexaose and maltoheptaose bound at the donor sites of cyclodextrin glycosyltransferase give insight into the mechanisms of transglycosylation activity and cyclodextrin size specificity.. Biochemistry, 39, 7772-7780. [PubMed id: 10869182] [DOI: 10.1021/bi000340x]

Date:

22-Mar-00

Release date:

22-Nov-00

Related entries:

1cdg
wild type enzyme
1cxi
wild type at 120 k
1cxf
e257q/d229n mutant at 120 k with a maltotetraose substrate
1cxk
e257q/d229n mutant at 120 k with a maltononaose substrate
1cxl
e257q/d229n mutant at 120 k with a covalently bound
... plus others (see Header records)

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Key reference

DOI no: 10.1021/bi000340x Biochemistry 39:7772-7780 (2000)

PubMed id: 10869182

Structures of maltohexaose and maltoheptaose bound at the donor sites of cyclodextrin glycosyltransferase give insight into the mechanisms of transglycosylation activity and cyclodextrin size specificity.

J.C.Uitdehaag, G.J.van Alebeek, B.A.van Der Veen, L.Dijkhuizen, B.W.Dijkstra.

ABSTRACT

The enzymes from the alpha-amylase family all share a similar alpha-retaining catalytic mechanism but can have different reaction and product specificities. One family member, cyclodextrin glycosyltransferase (CGTase), has an uncommonly high transglycosylation activity and is able to form cyclodextrins. We have determined the 2.0 and 2.5 A X-ray structures of E257A/D229A CGTase in complex with maltoheptaose and maltohexaose. Both sugars are bound at the donor subsites of the active site and the acceptor subsites are empty. These structures mimic a reaction stage in which a covalent enzyme-sugar intermediate awaits binding of an acceptor molecule. Comparison of these structures with CGTase-substrate and CGTase-product complexes reveals three different conformational states for the CGTase active site that are characterized by different orientations of the centrally located residue Tyr 195. In the maltoheptaose and maltohexaose-complexed conformation, CGTase hinders binding of an acceptor sugar at subsite +1, which suggests an induced-fit mechanism that could explain the transglycosylation activity of CGTase. In addition, the maltoheptaose and maltohexaose complexes give insight into the cyclodextrin size specificity of CGTases, since they precede alpha-cyclodextrin (six glucoses) and beta-cyclodextrin (seven glucoses) formation, respectively. Both ligands show conformational differences at specific sugar binding subsites, suggesting that these determine cyclodextrin product size specificity, which is confirmed by site-directed mutagenesis experiments.

Literature references that cite this PDB file's key reference

PubMed id Reference

17891389 Z.Li, M.Wang, F.Wang, Z.Gu, G.Du, J.Wu, and J.Chen (2007).
gamma-Cyclodextrin: a review on enzymatic production and applications.

Appl Microbiol Biotechnol, 77, 245-255.

16012834 K.Hirano, T.Ishihara, S.Ogasawara, H.Maeda, K.Abe, T.Nakajima, and Y.Yamagata (2006).
Molecular cloning and characterization of a novel gamma-CGTase from alkalophilic Bacillus sp.

Appl Microbiol Biotechnol, 70, 193-201.

14660599 K.S.Bak-Jensen, G.André, T.E.Gottschalk, G.Paës, V.Tran, and B.Svensson (2004).
Tyrosine 105 and threonine 212 at outermost substrate binding subsites -6 and +4 control substrate specificity, oligosaccharide cleavage patterns, and multiple binding modes of barley alpha-amylase 1.

J Biol Chem, 279, 10093-10102.

14993702 M.Akita, Y.Hatada, Y.Hidaka, Y.Ohta, M.Takada, Y.Nakagawa, K.Ogawa, T.Nakakuki, S.Ito, and K.Horikoshi (2004).
Crystallization and preliminary X-ray study of gamma-type cyclodextrin glycosyltransferase from Bacillus clarkii.

Acta Crystallogr D Biol Crystallogr, 60, 586-587.

12492486 H.Leemhuis, B.W.Dijkstra, and L.Dijkhuizen (2003).
Thermoanaerobacterium thermosulfurigenes cyclodextrin glycosyltransferase.

Eur J Biochem, 270, 155-162.

12423336 H.Mori, K.S.Bak-Jensen, and B.Svensson (2002).
Barley alpha-amylase Met53 situated at the high-affinity subsite -2 belongs to a substrate binding motif in the beta-->alpha loop 2 of the catalytic (beta/alpha)8-barrel and is critical for activity and substrate specificity.

Eur J Biochem, 269, 5377-5390.

12364331 L.K.Skov, O.Mirza, D.Sprogøe, I.Dar, M.Remaud-Simeon, C.Albenne, P.Monsan, and M.Gajhede (2002).
Oligosaccharide and sucrose complexes of amylosucrase. Structural implications for the polymerase activity.

J Biol Chem, 277, 47741-47747.

PDB codes: 1mvy 1mw0 1mw1 1mw2 1mw3

12196524 M.C.Abad, K.Binderup, J.Rios-Steiner, R.K.Arni, J.Preiss, and J.H.Geiger (2002).
The X-ray crystallographic structure of Escherichia coli branching enzyme.

J Biol Chem, 277, 42164-42170.

PDB code: 1m7x

11790748 N.Rashid, J.Cornista, S.Ezaki, T.Fukui, H.Atomi, and T.Imanaka (2002).
Characterization of an archaeal cyclodextrin glucanotransferase with a novel C-terminal domain.

J Bacteriol, 184, 777-784.

11288183 J.C.Uitdehaag, B.A.van der Veen, L.Dijkhuizen, R.Elber, and B.W.Dijkstra (2001).
Enzymatic circularization of a malto-octaose linear chain studied by stochastic reaction path calculations on cyclodextrin glycosyltransferase.

Proteins, 43, 327-335.

11282590 Y.Terada, H.Sanbe, T.Takaha, S.Kitahata, K.Koizumi, and S.Okada (2001).
Comparative study of the cyclization reactions of three bacterial cyclomaltodextrin glucanotransferases.

Appl Environ Microbiol, 67, 1453-1460.

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.