Crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis.

D. Sprogoe, L.A.M. van den Broek, O. Mirza, J.S. Kastrup, A.G.J. Voragen, M. Gajhede, L.K. Skov

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Abstract

Around 80 enzymes are implicated in the generic starch and sucrose pathways. One of these enzymes is sucrose phosphorylase, which reversibly catalyzes the conversion of sucrose and orthophosphate to d-Fructose and a-d-glucose 1-phosphate. Here, we present the crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis (BiSP) refined at 1.77 Å resolution. It represents the first 3D structure of a sucrose phosphorylase and is the first structure of a phosphate-dependent enzyme from the glycoside hydrolase family 13. The structure of BiSP is composed of the four domains A, B, B‘, and C. Domain A comprises the (ß/a)8-barrel common to family 13. The catalytic active-site residues (Asp192 and Glu232) are located at the tips of ß-sheets 4 and 5 in the (ß/a)8-barrel, as required for family 13 members. The topology of the B‘ domain disfavors oligosaccharide binding and reduces the size of the substrate access channel compared to other family 13 members, underlining the role of this domain in modulating the function of these enzymes. It is remarkable that the fold of the C domain is not observed in any other known hydrolases of family 13. BiSP was found as a homodimer in the crystal, and a dimer contact surface area of 960 Å2 per monomer was calculated. The majority of the interactions are confined to the two B domains, but interactions between the loop 8 regions of the two barrels are also observed. This results in a large cavity in the dimer, including the entrance to the two active sites.
Original languageEnglish
Pages (from-to)1156-1162
JournalBiochemistry
Volume43
Issue number5
DOIs
Publication statusPublished - 2004

Fingerprint

sucrose phosphorylase
Crystal structure
Catalytic Domain
Enzymes
Dimers
Sucrose
Phosphates
Glycoside Hydrolases
Hydrolases
Fructose
Oligosaccharides
Starch
Monomers
Topology
Crystals
Bifidobacterium adolescentis
Substrates

Keywords

  • neisseria-polysaccharea
  • glycosyl hydrolases
  • sequence alignment
  • maltogenic amylase
  • alpha-amylase
  • amylosucrase
  • crystallization
  • crystallography
  • neopullulanase
  • purification

Cite this

Sprogoe, D. ; van den Broek, L.A.M. ; Mirza, O. ; Kastrup, J.S. ; Voragen, A.G.J. ; Gajhede, M. ; Skov, L.K. / Crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis. In: Biochemistry. 2004 ; Vol. 43, No. 5. pp. 1156-1162.
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title = "Crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis.",
abstract = "Around 80 enzymes are implicated in the generic starch and sucrose pathways. One of these enzymes is sucrose phosphorylase, which reversibly catalyzes the conversion of sucrose and orthophosphate to d-Fructose and a-d-glucose 1-phosphate. Here, we present the crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis (BiSP) refined at 1.77 {\AA} resolution. It represents the first 3D structure of a sucrose phosphorylase and is the first structure of a phosphate-dependent enzyme from the glycoside hydrolase family 13. The structure of BiSP is composed of the four domains A, B, B‘, and C. Domain A comprises the ({\ss}/a)8-barrel common to family 13. The catalytic active-site residues (Asp192 and Glu232) are located at the tips of {\ss}-sheets 4 and 5 in the ({\ss}/a)8-barrel, as required for family 13 members. The topology of the B‘ domain disfavors oligosaccharide binding and reduces the size of the substrate access channel compared to other family 13 members, underlining the role of this domain in modulating the function of these enzymes. It is remarkable that the fold of the C domain is not observed in any other known hydrolases of family 13. BiSP was found as a homodimer in the crystal, and a dimer contact surface area of 960 {\AA}2 per monomer was calculated. The majority of the interactions are confined to the two B domains, but interactions between the loop 8 regions of the two barrels are also observed. This results in a large cavity in the dimer, including the entrance to the two active sites.",
keywords = "neisseria-polysaccharea, glycosyl hydrolases, sequence alignment, maltogenic amylase, alpha-amylase, amylosucrase, crystallization, crystallography, neopullulanase, purification",
author = "D. Sprogoe and {van den Broek}, L.A.M. and O. Mirza and J.S. Kastrup and A.G.J. Voragen and M. Gajhede and L.K. Skov",
year = "2004",
doi = "10.1021/bi0356395",
language = "English",
volume = "43",
pages = "1156--1162",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "5",

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Crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis. / Sprogoe, D.; van den Broek, L.A.M.; Mirza, O.; Kastrup, J.S.; Voragen, A.G.J.; Gajhede, M.; Skov, L.K.

In: Biochemistry, Vol. 43, No. 5, 2004, p. 1156-1162.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis.

AU - Sprogoe, D.

AU - van den Broek, L.A.M.

AU - Mirza, O.

AU - Kastrup, J.S.

AU - Voragen, A.G.J.

AU - Gajhede, M.

AU - Skov, L.K.

PY - 2004

Y1 - 2004

N2 - Around 80 enzymes are implicated in the generic starch and sucrose pathways. One of these enzymes is sucrose phosphorylase, which reversibly catalyzes the conversion of sucrose and orthophosphate to d-Fructose and a-d-glucose 1-phosphate. Here, we present the crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis (BiSP) refined at 1.77 Å resolution. It represents the first 3D structure of a sucrose phosphorylase and is the first structure of a phosphate-dependent enzyme from the glycoside hydrolase family 13. The structure of BiSP is composed of the four domains A, B, B‘, and C. Domain A comprises the (ß/a)8-barrel common to family 13. The catalytic active-site residues (Asp192 and Glu232) are located at the tips of ß-sheets 4 and 5 in the (ß/a)8-barrel, as required for family 13 members. The topology of the B‘ domain disfavors oligosaccharide binding and reduces the size of the substrate access channel compared to other family 13 members, underlining the role of this domain in modulating the function of these enzymes. It is remarkable that the fold of the C domain is not observed in any other known hydrolases of family 13. BiSP was found as a homodimer in the crystal, and a dimer contact surface area of 960 Å2 per monomer was calculated. The majority of the interactions are confined to the two B domains, but interactions between the loop 8 regions of the two barrels are also observed. This results in a large cavity in the dimer, including the entrance to the two active sites.

AB - Around 80 enzymes are implicated in the generic starch and sucrose pathways. One of these enzymes is sucrose phosphorylase, which reversibly catalyzes the conversion of sucrose and orthophosphate to d-Fructose and a-d-glucose 1-phosphate. Here, we present the crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis (BiSP) refined at 1.77 Å resolution. It represents the first 3D structure of a sucrose phosphorylase and is the first structure of a phosphate-dependent enzyme from the glycoside hydrolase family 13. The structure of BiSP is composed of the four domains A, B, B‘, and C. Domain A comprises the (ß/a)8-barrel common to family 13. The catalytic active-site residues (Asp192 and Glu232) are located at the tips of ß-sheets 4 and 5 in the (ß/a)8-barrel, as required for family 13 members. The topology of the B‘ domain disfavors oligosaccharide binding and reduces the size of the substrate access channel compared to other family 13 members, underlining the role of this domain in modulating the function of these enzymes. It is remarkable that the fold of the C domain is not observed in any other known hydrolases of family 13. BiSP was found as a homodimer in the crystal, and a dimer contact surface area of 960 Å2 per monomer was calculated. The majority of the interactions are confined to the two B domains, but interactions between the loop 8 regions of the two barrels are also observed. This results in a large cavity in the dimer, including the entrance to the two active sites.

KW - neisseria-polysaccharea

KW - glycosyl hydrolases

KW - sequence alignment

KW - maltogenic amylase

KW - alpha-amylase

KW - amylosucrase

KW - crystallization

KW - crystallography

KW - neopullulanase

KW - purification

U2 - 10.1021/bi0356395

DO - 10.1021/bi0356395

M3 - Article

VL - 43

SP - 1156

EP - 1162

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 5

ER -